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ENVIRONMENTAL RISK MANAGEMENT AUTHORITY
DECISION
Amended under s67A on 23 August 2007
Dated signed: 31 July 2002
GMC02006
To import into containment organisms under section 40(1)(a) of
the Hazardous Substances and New Organisms (HSNO) Act.
University of Otago
Applicant
To import into containment genetically modified mice to investigate
Purpose
the molecular and cellular properties of the gonatrophin-releasing
hormone (GnRH) neurons that are responsible for regulating fertility in
all mammalian species
13 June 2002
Date received
Consideration period 05 – 18 July 2002
The Genetically Modified Organism Standing Committee of the
Considered by
Environmental Risk Management Authority (the Authority).
Application code
Application type
1
Summary of Decision
The application to import into containment 13 strains of genetically modified (GM) mice (as
described in Annex 2 of this decision) is approved, with controls (as listed in Annex 1 of this
decision), having been considered in accordance with the relevant provisions of the
Hazardous Substances and New Organisms (HSNO) Act 1996 and the Hazardous Substances
and New Organisms (Methodology) Order 1998 (the Methodology).
In accordance with section 45(3) of the HSNO Act, the Committee must give reasons for the
decision in writing. These reasons are summarised in the following sections.
2
Summary of consideration process & relevant
legislative criteria
2.1
Decision-making Committee
The application was considered by the Genetically Modified Organism Standing Committee
of the Authority, appointed in accordance with section 19(2)(b) of the HSNO Act. The
Committee comprised the following members: Mrs Jane Lancaster (Chair), Mrs Jill White
and Mr George Clark.
2.2
Consideration approach and sequence
The decision was determined in accordance with section 45 of the HSNO Act, taking into
account additional matters to be considered under section 44, and matters relevant to the
purpose of the HSNO Act (as specified under Part II of the Act). Unless otherwise stated,
references to section numbers in this decision refer to sections of the HSNO Act.
Consideration of the application followed the relevant provisions of the Hazardous
Substances and New Organisms (Methodology) Order 1998 (the Methodology), as specified
in more detail below. Unless otherwise stated, references to clause numbers in this decision
refer to clauses of the Methodology.
In accordance with clause 24 of the Methodology, the approach to consideration adopted by
the Committee was to look sequentially at identification, assessment and evaluation of risks,
costs and benefits. The terms adverse effects and beneficial effects are used in this decision
document to incorporate the terms ‘risks, costs and benefits’. Adverse effects are defined as
including risks and associated costs. Costs are defined as values of negative effects (expressed
in monetary or non-monetary terms). Beneficial effects are defined as the value of positive
effects (expressed in monetary or non-monetary terms). Qualitative scales used by the
Committee to measure likelihood and magnitude of effect, are provided as Annex 3 of this
decision.
Adverse and beneficial effects have been categorised in terms of their area of impact on:

Environment

Māori culture

Economy

Human health and safety

Society and community
In assessing adverse effects, issues affecting the adequacy of the containment regime and
potential for population establishment and population eradication were considered (as
required by sections 37 and 44 of the HSNO Act and clause 10(e) of the Methodology). The
containment regime was considered in the context of a risk management regime for
controlling the identified risks (clauses 12(d) and 24). In doing so, the Committee set controls
to satisfactorily provide for the matters specified in the Third Schedule (Part I) of the HSNO
Act. It was then considered whether or not there were any residual risks that required further
consideration.
Beneficial effects associated with the application were considered in accordance with the
Methodology clauses 9, 10, 13, and 14 and section 6(e) of the HSNO Act.
The decision-making process is described in more detail in the following sections, with
further reference to legislative requirements under the HSNO Act and Methodology. The
legislative criteria taken into account in the overall evaluation are summarised in section 5 of
this decision.
2.3
Application receipt
The application was formally received by ERMA New Zealand on 13 June 2002 pursuant to
section pursuant to section 40(1)(a) of the HSNO Act. ERMA New Zealand verified that the
application contained adequate information to be processed on 18 June 2002.
Environmental Risk Management Authority Decision: Application GMC02006
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Purpose of the application
The purpose of the application is to import into containment genetically modified (transgenic
and "knockout") mice (as specified in Annex 2) to investigate the molecular and cellular
properties of the gonadotrophin-releasing hormone (GnRH) neurones that are responsible for
regulating fertility in all mammalian species. In accordance with section 45(1)(a)(i) of the
HSNO Act, the Committee determined that the purpose was appropriate under section
39(1)(h): Such other purposes as the Authority thinks fit.
As noted in section 8 of application, the applicant intends to use some of the GM mice to
maintain strains by backcross, or to breed with other transgenic strains to enhance desired
characteristics. The Committee notes that backcrossing to the parental strain does not
constitute development of a genetically modified organism (GMO) and hence does not require
a separate approval under the HSNO Act. However, crossing different strains of GM mice
constitutes the development of a new GMO, and would therefore require the applicant to
submit an application to develop a GMO under the HSNO Act.
Discretion to publicly notify application receipt
The Authority has discretion as to whether or not receipt of an application to import into
containment any new organism is publicly notified. In this case the application was not
publicly notified (following the ERMA New Zealand Corporate Manual Item 3.2.23), as the
organisms to be imported would be defined as low risk (Category A or B) in the HSNO (LowRisk Genetic Modification) Regulations 1998.
Consultation with departments and Crown entities
In accordance with clauses 2(2)(e) and 5 of the Methodology and section 58(c) of the HSNO
Act, departments and Crown entities which were likely to have an interest in the application
were notified of application receipt. As such, the Department of Conservation (DoC) and
Ministry of Agriculture and Forestry (MAF) were invited to comment on this application. It is
noted that DoC does not routinely provide comment on GM mouse applications (by prior
arrangement with ERMA New Zealand) unless particularly novel issues are raised. This is
due to the large number of GM mouse applications that have already been processed by
ERMA New Zealand that present issues of the same (or very similar) nature. Neither DoC nor
MAF Biosecurity Authority had any specific comments on this application.
2.4
Information available for consideration
The documents available for the consideration included the application and published
references as cited in the application. In addition, staff advice was provided to the Committee
in the form of an Evaluation and Review (E&R) Report. The aim of the E&R Report is to
assist and support the Committee’s decision-making by consolidating information provided
by the applicant, and obtained from other sources, into a format and sequence that is
consistent with the decision-making requirements of the HSNO Act and the Methodology.
Recognised techniques were used in identifying, assessing, and evaluating the relevant
information, as required under clause 24 of the Methodology. Techniques for identifying and
preparing information on risks, costs and benefits were based on internal procedures as
specified in the ERMA New Zealand Technical Guide publications. The information was
evaluated to provide an opinion on its quality and credibility, as well as to identify key issues
associated with the application.
Environmental Risk Management Authority Decision: Application GMC02006
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3
Identification and assessment of adverse and beneficial
effects
3.1
Assessment of containment regime adequacy
In assessing adverse effects, the Committee considered issues affecting the adequacy of the
containment regime (in accordance with section 45(i)(a)(iii) of the HSNO Act); the potential
for population establishment and population eradication (sections 37 and 44 of the HSNO Act
and clause 10(e) of the Methodology); and other matters in order to give effect to the purpose
of the HSNO Act (section 45(2)). Risk management techniques were assessed in relation to
the identified risks (clauses 12(d) and 24). As such, the assessment of adverse effects (refer to
section 4 of this decision) was taken into account in setting the containment requirements that
are discussed in this section.
In assessing issues affecting the adequacy of the containment regime, the Committee concurs
with the identification and assessment of these issues provided in sections 3 and 4 of the E&R
Report. Key issues are summarised below.
Ability to escape from containment
The controls imposed by this approval (as specified in Annex 1) address the matters detailed
in the Third Schedule Part I: Matters to be addressed by containment controls for importing,
developing or field testing of genetically modified organisms of the HSNO Act. These
controls incorporate requirements for management of risks (under clauses 12(d) and 24 of the
Methodology) posed by the GM mice subject to this approval. The controls have been
imposed to ensure that exposure of laboratory workers and other persons, and the outside
environment, to risks posed by the organisms are negligible.
The basis of the containment regime is that the GM mice shall be held in a containment
facility that is registered and operated in accordance with the MAF Biosecurity
Authority/ERMA New Zealand Standard 154.03.03 (Containment Facilities for Vertebrates).
General containment requirements for safe management of vertebrates in the laboratory
environment are addressed in this standard, with cross-reference to detailed physical
containment (PC) requirements in the Australian New Zealand Standard AS/NZS
2243.3:2002 Safety in Laboratories Part 3: Microbiological aspects and containment
facilities. The physical containment level has been set at PC2, as per the AS/NZ standard
(refer to Control 1.2, Annex 1). The other controls specified in Annex 1 emphasize and/or
clarify how the above containment standards address the matters required under the Third
Schedule (Part I) of the HSNO Act.
As with previous similar applications for GM mice (refer to section 4 of this decision), the
Committee considers that no additional requirements (over and above the Vertebrate Standard
154.03.03) are warranted to manage risks associated with this application.
The Committee notes that the University of Otago intends to hold the mice at three associated
facilities (ie the Wellington School of Medicine; the Christchurch School of Medicine and the
University of Otago, Dunedin campus). These three facilities are currently registered as
containment facilities for vertebrate laboratory animals under MAF/ERMA standard
(154.03.03) at PC2.
Environmental Risk Management Authority Decision: Application GMC02006
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In assessing the ability of the GM mice to escape from containment, the Committee concludes
that the accidental or deliberate removal of the GM mice from containment would be very
unlikely, based on the containment regime imposed and the experience of the staff in the
facilities involved.
Ability of the organisms to establish undesirable self-sustaining populations and ease of
eradication
In accordance with sections 44 and 37 of the HSNO Act the Committee considered the ability
of the organisms to establish undesirable self-sustaining populations, should they escape from
containment, and the ease with which such populations could be eradicated. In evaluating
these matters the Committee took into account the nature of the organisms.
The Committee noted that the GM mice are in-bred laboratory strains, and would therefore be
poorly adapted for survival in the wild. As such in the very unlikely event that a breeding pair
of genetically modified mice escaped from containment, they are very unlikely to form a
distinct, self-sustaining population.
If the GM mice escaped and bred with wild endemic mice, the distinctive genetic traits of the
GM strains may or may not be lost during subsequent breeding, depending on the viability
and mating success of the mice containing them. In addition, some genetic modifications are
spontaneously lost or do not express themselves (for a variety of reasons) so that the
modifications may not be retained or expressed even if it was not deleterious. There is
uncertainty over whether some of the traits would remain in the population, but if there is no
selective advantage then they would be at a low frequency and may eventually be lost.
Anecdotal reports from those handling laboratory mice include mention that when such mice
are out of their cages they do not tend to run away, so can be easily captured. However, in the
unlikely event that the mice escaped the containment facility into the uncontrolled
environment, the Committee considers it is unlikely that they could be recovered. While it is
generally accepted that mice bred and raised in sheltered conditions are very unlikely to
survive in the external environment for longer than a few days, there is a minor degree of
uncertainty in this contention. However, given the containment controls imposed, and the
biological characteristics of the GM mice, it is very unlikely that self-sustaining populations
of the GM mice would establish should an escape from containment occur. The Committee
thus considers that eradication of these organisms would not be necessary.
Other matters
The Committee considered what other matters might be relevant in setting controls to provide
for matters outside the Third Schedule, in order to give effect to the purpose of the HSNO Act
(in accordance with section 45(2)).
The Committee considers that it is inappropriate to impose a restricted time limit on this
approval because the approval is likely to be used over a period of time. The Committee also
notes that the controls imposed within this approval have been used in a large number of
previous approvals, and as such do not consider that it is necessary for the applicant to report
to ERMA New Zealand specifically on the adequacy of the controls imposed by this decision.
In addition, the Committee considers that additional controls to measure or monitor for
adverse effects are not necessary in this case since the work will be conducted within standard
containment facilities, and there is no intention to field test or release the organisms and there
will be no development of novel viruses.
Environmental Risk Management Authority Decision: Application GMC02006
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In conclusion, no additional matters were identified by the Committee to be relevant in order
to give effect to the purpose of the HSNO Act.
Summary of assessment of containment regime adequacy
The Committee considers that with the containment controls it has imposed, as discussed
above and detailed in Annex 1 of this decision, it is very likely that the GM mice can be
adequately contained. The basis of the containment regime is the requirement for the
containment facility to be registered and operate in accordance with the MAF Biosecurity
Authority/ERMA New Zealand Standard 154.03.03 (Containment Facilities for Vertebrates).
No additional requirements are considered necessary in order to manage risks associated with
the application. In the unlikely event that the GM mice escape or are removed from
containment, it is very unlikely that self-sustaining populations would establish in the external
environment. The possible impacts of such a breach are considered to be minimal to minor in
terms of adverse effects (refer to section 3.2 below, for more detail regarding adverse effects).
3.2
Identification and assessment of adverse effects
The Committee identified and assessed adverse effects (ie risks and costs) related to the
application in accordance with the legislative criteria described in section 2.2 above. The
Committee concurs with the identification and assessment of adverse effects provided in
sections 2, 3 and 5 of the E&R Report. The Committee notes that no adverse economic, social
or community effects were identified. Therefore no specific assessment of these matters was
conducted. Key issues relating to identification and assessment of adverse effects are
summarised below.
Characteristics of organism
In assessing adverse effects, the Committee considered the characteristics of the organisms
and the genetic modifications. The Committee notes the assessment of organism
characteristics provided in section 2 of the E&R Report, including that such inbred mouse
strains (in this case with artificial modifications to their genomes) are commonly used to
develop experimental models. Furthermore, if the GM mice were developed in New Zealand,
they would meet the low risk criteria (category B(b)(ii)B)) as defined in the HSNO (Low-Risk
Genetic Modification) Regulations 1998, and could therefore be processed via the rapid
assessment route.
Associated organisms
In assessing adverse effects, the Committee considered the nature of organisms that might be
associated with the GM mice. The Committee concurs with the assessment provided in
section 5 of the E&R Report, that any pathogen or parasite associated with the GM mice (ie
separable or inseparable) is unlikely to pass undetected through the quarantine regime
required by MAF (in accordance with an Import Health Standard issued under the Biosecurity
Act). Based on the characteristics of the laboratory-bred GM mice, it is very unlikely for any
inseparable organisms of significance (ie pathogens or parasites) to be inadvertently imported.
Environmental Risk Management Authority Decision: Application GMC02006
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Adverse environmental effects
In the unlikely event that the GM mice escaped and bred with endemic mice, the distinctive
traits of the GM strains would very likely be lost through cross-breeding as the retention of
these distinctive traits generally requires selective inbreeding. The Committee considers that
since the modifications do not produce toxins or infectious particles, or increase the
reproductive or escape ability of the mice, it is very unlikely that the GM mice could exert
any adverse environmental effect, in the unlikely event of their escape from containment. The
Committee considers that the magnitude of any adverse effects on native or valued flora and
fauna would be minimal or minor, given that survival of the mice is likely to be compromised
and the modifications are very unlikely to have adverse effects on other organisms.
Adverse human health and safety effects
The Committee considers that the magnitude of any adverse effects on human health and
safety are likely to be minimal, given the disease-free status of the animals, that they will be
handled by only a small number of authorised research staff, and that the mice are very
unlikely to survive without human intervention if they escape into the uncontrolled
environment.
Adverse Māori cultural effects
From the information provided, the Committee considers that the application poses no risk to
the relationship between Māori culture and their traditions with their ancestral lands, water,
sites, waahi tapu, valued flora and fauna and other taonga. Therefore, it was appropriate that
the applicant did not consult with Māori regarding this application.
Costs
The Committee concurs with the conclusion drawn in section 3 of the E&R Report that any
costs associated with the application are covered as factors of adverse effects. No other costs
have been identified. Costs of the application will be borne by the applicant. No costs are
likely to be imposed on third parties by approval of the application. Therefore, no specific
assessment of costs was conducted.
Summary of assessment of adverse effects
The Committee considers that there are no significant adverse effects associated with this
application to import into containment of GM mice. In reaching this conclusion, the adequacy
of the containment regime has been taken into account, as well as the potential for population
establishment and population eradication if the mice escape containment. The Committee is
satisfied that the controls provide for the matters specified in the Third Schedule (Part I) of
the HSNO Act. No residual risks have been identified that require further consideration. The
Committee therefore concludes that the adverse effects associated with this application are
negligible. As such, the Committee similarly considers the specific aspects of risk defined
under clause 33 are not significant.
3.3
Identification and assessment of beneficial effects
Benefits that may arise from any of the matters set out in clauses 9 and 10 of the Methodology
were considered in terms of clause 13. The Committee concurs with the identification and
assessment of beneficial effects provided in sections 3 and 5 of the E&R Report.
Environmental Risk Management Authority Decision: Application GMC02006
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Under clause 13(a), the Committee considered the primary benefit associated with this
application to be increased scientific knowledge (non-monetary) gained through investigating
the molecular and cellular properties of gonadotrophin-releasing hormone (GnRH) neurones.
The Committee notes that while increased scientific knowledge likely, the magnitude and
expected value of beneficial effects (clause 13(b)) is uncertain. Immediate benefits are likely
to accrue directly to the applicant (clause 13(c)).
Indirect and/or longer-term benefits for human health and the economy are possible, but
highly uncertain in terms of their magnitude of effect. This will depend on whether or not the
results of research are suitable to translate into practical outcomes. The Committee considers
that economic benefits relating to the applicant organisation’s ability to attract funds
(domestically and internationally) are realistic, as a consequence of the research associated
with this application, to support the progress of future research.
4
Relationship with previous approvals
In considering application GMC02006, the Committee has reflected on previous decisions
that involve similar issues to those raised by this application. However, the Committee notes
that each application is considered on its merits, and therefore the outcome of the current
application is not bound by the stance taken in previous decisions.
As noted in section 4 of the application, the University of Otago currently has HSNO Act
approvals to import, breed and develop many GM mouse strains in containment for
experimental research at PC2 (for their associated facilities at the Wellington School of
Medicine; the Christchurch School of Medicine and the University of Otago, Dunedin
campus). The Committee considers that the risks posed by this application cannot be
distinguished from those identified in relation to several previous GM mouse applications,
including application GMC02003 (Malaghan Institute of Medical Research) and University of
Otago applications GMC00009, GMC00015, GMC00019, GMC99005 and GMC01006.
These imports involved genetic modifications that were also considered to present negligible
risks. The containment regime for all these approvals require Containment Facilities for
Vertebrate Laboratory Animals (MAF Biosecurity Authority/ERMA New Zealand Standard
154.03.03), at Physical Containment Level 2 (PC2).
5
Overall evaluation of adverse and beneficial effects
In reaching its decision on this application, the Committee records that the following criteria
in the HSNO Act and Methodology have been particularly relied on (in accordance with
clauses 21 and 36(2)(b) of the Methodology):

The application has been considered in the context of the purpose and principles of the
HSNO Act (sections 4 – 8 inclusive).

Pursuant to section 45(1)(a)(i) of the HSNO Act, the Committee is satisfied that this
that the purpose of the application is appropriate under section 39(1)(h): Such other
purposes as the Authority thinks fit.

The information provided by the applicant is relevant and appropriate to the scale and
significance of the risks and costs associated with the application (clause 8 of the
Methodology).
Environmental Risk Management Authority Decision: Application GMC02006
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6
The overall evaluation of risks, costs and benefits was carried out having regard to
Clauses 22 and 34 of the Methodology and in accordance with the tests in clause 26 of
the Methodology and section 45 of the Act.
The Committee is satisfied that the modified mice can be adequately contained
(sections 45(1)(a)(iii) and 44(b) of the HSNO Act), under the controls required by this
decision (refer to Annex 1). In relation to the additional matters to be considered under
section 37 of the HSNO Act, the Committee considers that it is very unlikely for the
genetically modified mice to escape or be removed inadvertently from containment
and form a self-sustaining population, and that eradication would be unnecessary
given that the organisms would be unlikely to survive outside of containment.
The Committee did not identify any potential significant risks to the environment, to
public health and safety, or to Māori and their taonga associated with this application.
In accordance with section 45 of the HSNO Act and clauses 9, 10 and 12 of the
Methodology, the Committee considers that to a high level of certainty, both the
magnitude and probability of occurrence of adverse effects arising from importation of
the genetically modified mice subject to application GMC02006 is negligible.
In accordance with clause 33 of the Methodology, the Authority must have regard to
the extent to which particular risk characteristics exist. In concluding that the adverse
effects associated with this application are negligible, the Committee similarly
considers the specific aspects of risk defined under clause 33 are not significant.
The Committee has reached its decision on this application under clause 26 of the
Methodology, and considers that the benefits associated with the importation and use
of the GM mice in containment outweigh the costs. In reaching this conclusion, the
Committee considered all the possible beneficial and adverse effects of the organisms
in accordance with sections 45(1)(a)(ii) and (iii) of the HSNO Act.
Decision
The application to import into containment genetically modified mice (as described in Annex
2 of this decision) is approved in accordance with section 45 of the HSNO Act. As required
under section 45(2) the approval is subject to controls (as listed in Annex 1 of this decision).
Mrs Jane Lancaster
Date: 31 July 2002
Chair, GMO New Organisms Standing Committee of the Authority
Amendment: November 2006
Changes to controls:
 Addition of footnotes to the containment facility references and the Australian/New
Zealand containment facility references to “future proof” the decision
 Standardise the wording of the breach of containment control
 Removal of the control regarding inspection of facilities by the Authority, its agent or
enforcement officers
____________________________
Dr Kieran Elborough
Chair, GMO Standing Committee
Date: 23 August 2007
Environmental Risk Management Authority Decision: Application GMC02006
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Annex 1: Controls required by this approval
In order to satisfactorily address the matters detailed in the Third Schedule Part I:
Containment controls for importing, developing or field testing of genetically modified
organisms1 of the HSNO Act, and other matters in order to give effect to the purpose of the
HSNO Act (section 45(2)), the Authority’s approval of this application is subject to the
following controls:
1.
To limit the likelihood of any accidental release of any organism or any viable
genetic material2:
The person responsible for a particular research area and/or the person responsible for the
operation of the containment facility shall inform all personnel involved in the handling of the
organisms of the Authority’s controls.
The containment facility in which the organisms are maintained shall be in accordance
with the MAF Biosecurity Authority/ERMA New Zealand Standard 154.03.033. Containment
Facilities for Vertebrate Laboratory Animals, at Physical Containment Level 2 (PC2) as
defined in AS/NZS Standard 2243.3.20023. Safety in Laboratories Part 3: Microbiological
Aspects and Containment Facilities.
The construction and operation of the containment facilities (‘the facility’) in which the
organisms are maintained, shall be in accordance with the relevant standards listed in control
1.2 above.
2.
To exclude unauthorised people from the facility:
The identification of entrances, numbers of and access to entrances, and security
requirements for the entrances and the facility shall be in compliance with the standards listed
in control 1.2.
3.
To exclude other organisms from the facility and to control undesirable and unwanted
organisms within the facility:
Construction and operation of the containment facility shall comply with the requirements
of the standards listed in control 1.2 relating to the exclusion of other organisms from the
facility and the control of undesirable and unwanted organisms within the facility.
4.
To prevent unintended release of the organism by experimenters working with
the organism:
Construction and operation of the containment facility shall comply with the requirements
of the standards listed in control 1.2 relating to the prevention of unintended release of the
organisms by experimenters working with the organisms.
Bold headings refer to Matters to be Addressed by Containment Controls for Development and Field Testing of Genetically
Modified Organisms, specified in the Third Schedule of the HSNO Act 1996.
1
Viable Genetic Material is biological material that can be resuscitated to grow into tissues or organisms. It can be
defined to mean biological material capable of growth even though resuscitation procedures may be required, eg
when organisms or parts thereof are sublethally damaged by being frozen, dried, heated, or affected by chemical.
2
Any reference to this standard in these controls refers to any subsequent version approved or endorsed by ERMA
New Zealand
3
Environmental Risk Management Authority Decision: Application GMC02006
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5.
To control the effects of any accidental release or escape of an organism:
Construction and operation of the containment facility shall comply with the requirements
of the standards listed in control 1.2 relating to controlling the effects of any accidental
release or escape of an organism.
If a breach of containment occurs, the facility operator must ensure that the MAF
Inspector responsible for supervision of the facility has received notification of the breach
within 24 hours.
In the event of any breach of containment of the organisms, the contingency plan for the
attempted retrieval or destruction of any viable material of the organism that has escaped shall
be implemented immediately. The contingency plan shall be included in the containment
manual in accordance with the requirements of standards listed in control 1.2.
6.
Inspection and monitoring requirements for containment facilities:
The operation of the containment facilities shall comply with the requirements contained
in the standards listed in control 1.2 relating to the inspection and monitoring requirements for
containment facilities.
The containment manual shall be updated, as necessary, to address the implementation of
the controls imposed by this approval, in accordance with the Standards listed in control 1.2.
7.
Qualifications required of the persons responsible for implementing those
controls:
The training of personnel working in the facility shall be in compliance with the standards
listed in control 1.2.
Environmental Risk Management Authority Decision: Application GMC02006
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Annex 2: Organism descriptions
The 13 genetic modifications to mouse strains C57Bl6/J and C57Bl6/J x CBA/Ca have each
been assigned an organism name (ie a unique identification) and description for the ERMA
New Zealand Organism Register, as follows:
1.
Mus musculus (Linnaeus 1758) strains C57Bl6/J and C57Bl6/J x CBA/Ca,
GnRH-GFP (GMC02006)
Organism description:
Laboratory mice with integrated expression cassettes containing variants of the enhanced
green fluorescent protein (eGFP) gene under the control of normal and truncated versions of
the mouse gonadotropin-releasing hormone (GnRH) promoter. Normal or truncated versions
of the GnRH promoter drives the expression of variants of eGFP.
Modification introduced: An expression cassette containing variable amounts of the mouse
GnRH promoter driving the jellyfish (Aequorea victoria) green fluorescent protein gene is
integrated. The GFP reporters are modified by different point mutations to enhance brightness
and/or reduce toxicity.
Phenotype: No obvious changes in phenotype.
Detection of genetically modified animals: Primarily by genomic PCR using primers which
detect the eGFP gene.
References: Spergel DJ, Kruth U, Hanley DF, Sprengel R, Seeburg PH. (1999) GABA-and
glutamate-activated channels in green fluorescent protein-tagged gonadotropin-releasing
hormone neurone in transgenic mice. J. Neurosci. 19, 2037-2050.
Other names: Mus musculus (animal, vertebrate, mammal, mouse, GM, mice)
2.
Mus musculus (Linnaeus 1758) strains C57Bl6/J and C57Bl6/J x CBA/Ca,
GnRH-LacZ (GMC02006)
Organism description:
Laboratory mice with integrated expression cassettes containing the lacZ reporter gene under
the control of normal and truncated versions of the mouse gonadotropin-releasing hormone
(GnRH) promoter. Normal and truncated versions of the GnRH promoter drives the
expression of LacZ.
Modification introduced: An expression cassette containing variable amounts of the mouse
GnRH promoter and the lacZ gene is integrated.
Phenotype: No obvious changes in phenotype.
Detection of genetically modified animals: Primarily by genomic PCR using primers which
detect the LacZ gene. An antibody recognising beta-galactosidase, the protein product of
LacZ, can also be used to detect the transgene in GnRH neurones.
References: Pape J-R, Skynner MJ, Allen ND, Herbison AE. (1999) Transgenics identify
distal 5'- and 3' sequences specifying gonadotropin-releasing hormone expression in adult
mice. Mol. Endocrinol. 13, 2203-2211.
Other names: Mus musculus (animal, vertebrate, mammal, mouse, GM, mice)
Environmental Risk Management Authority Decision: Application GMC02006
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3.
Mus musculus (Linnaeus 1758) strains C57Bl6/J and C57Bl6/J x CBA/Ca,
GnRH-NR (GMC02006)
Organism description:
Laboratory mice with integrated expression cassettes containing the nitroreductase (NR) gene
under the control of normal and truncated versions of the mouse gonadotropin-releasing
hormone (GnRH) promoter. The GnRH promoter drives the expression of nitroreductase.
Modification introduced: An expression cassette containing 5kb of the mouse GnRH
promoter and the E.coli nitroreductase gene is integrated.
Phenotype: The females in one line (GNR2300) exhibit reduced fertility whereas the other
mice exhibit no obvious changes in phenotype.
Detection of genetically modified animals: Primarily by genomic PCR using primers which
detect the nitroreductase gene.
References: (1) Gamble JA, Pape JR, Allen JP, Skynner MJ, Bicknell RJ, Herbison AE
(2001) Disrupted GnRH neuronal migration in the GNR2300 mouse line. Soc. Neurosci.
Abstracts 27, 731.1 (2) Isles AR, Ma D, Milsom C, Skynner MJ, Cui W, Clark J, Keverne EB,
Allen ND (2001) Conditional ablation of neurones in transgenic mice. J Neurobiol, 47, 183193.
Other names: Mus musculus (animal, vertebrate, mammal, mouse, GM, mice)
4.
Mus musculus (Linnaeus 1758) strains C57Bl6/J and C57Bl6/J x CBA/Ca,
GnRH-Cre (GMC02006)
Organism description:
Laboratory mice with an integrated expression cassette containing the Cre recombinase gene
under the control of the mouse gonadotropin-releasing hormone (GnRH) promoter. The
GnRH promoter drives the expression of Cre recombinase.
Modification introduced: An expression cassette containing 12kb of the mouse GnRH locus
and the Cre recombinase gene is integrated. The Cre recombinase is originally dervied from
bacteriophage P1.
Phenotype: The mice exhibit no obvious changes in phenotype.
Detection of genetically modified animals: Primarily by genomic PCR using primers
specific to the incorporated expression cassette. An antibody specific to the Cre recombinase
is commercially available and can be used to detect Cre proteins in GnRH neurones.
References: Tsien JZ, Chen DF, Gerber D, Tom C, Mercer EH, Anderson DJ, Mayford, M,
Kandel ER, Tonegawa S (1996) Sub region- and cell type-restricted gene knockout in mouse
brain. Cell 87, 1317-1326. (Mice made in the Herbison laboratory, The Babraham Institute,
Cambridge, UK but are not yet published).
Other names: Mus musculus (animal, vertebrate, mammal, mouse, GM, mice)
5.
Mus musculus (Linnaeus 1758) strains C57Bl6/J and C57Bl6/J x CBA/Ca,
GnRH-pericam (GMC02006)
Organism description:
Laboratory mice with integrated expression cassettes containing variants of the “pericam
gene” under the control of the mouse gonadotropin-releasing hormone (GnRH) promoter. The
GnRH promoter drives the expression of circularly permuted green fluorescent protein
(Pericam).
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Modification introduced: An expression cassette containing 12kb of the mouse GnRH locus
and a circularly permuted (ie the beginning and end of the gene have been switched) jellyfish
(Aequorea victoria) green fluorescent protein gene to calmodulin (Pericam) is integrated.
Phenotype: The mice exhibit no obvious changes in phenotype.
Detection of genetically modified animals: Primarily by genomic PCR using primers
specific to the incorporated expression cassette.
References: Nagai T, Sawano A, Park ES, Miyawaki A. (2001) Circularly permuted green
fluorescent proteins engineered to sense Ca2+. Proc. Nat. Acad. Sci. 98, 3197-3202. (Mice
made in the Herbison laboratory, The Babraham Institute, Cambridge, UK but are not yet
published).
Other names: Mus musculus (animal, vertebrate, mammal, mouse, GM, mice)
6.
Mus musculus (Linnaeus 1758) strains C57Bl6/J and C57Bl6/J x CBA/Ca,
TH9-LacZ (GMC02006)
Organism description:
Laboratory mice with an integrated expression cassette containing the lacZ reporter gene
under the control of the rat tyrosine hydroxylase (TH) promoter. The TH promoter drives the
expression of LacZ.
Modification introduced: An expression cassette containing 9kb of rat TH promoter and the
LacZ gene is integrated.
Phenotype: No obvious changes in phenotype.
Detection of genetically modified animals: Primarily by genomic PCR using primers which
detect the LacZ gene. An antibody recognising beta-galactosidase, the protein product of
LacZ, can also be used to detect the transgene in TH-expressing neurones.
References: Min N, Joh TH, Kim KS, Peng C, Son JH. (1994) 5' upstream DNA sequence of
the rat tyrosine hydroxylase gene directs high-level and tissue-specific expression to
catecholaminergic neurons in the central nervous system of transgenic mice. Mol. Brain Res.
27, 281-289.
Other names: Mus musculus (animal, vertebrate, mammal, mouse, GM, mice)
7.
Mus musculus (Linnaeus 1758) strains C57Bl6/J and C57Bl6/J x CBA/Ca,
TH-GFP (GMC02006)
Organism description:
Laboratory mice with an integrated expression cassette containing an enhanced green
fluorescent protein (eGFP) gene under the control of the tyrosine hydroxylase (TH) promoter.
The TH promoter drives the expression of eGFP.
Modification introduced: An expression cassette containing the rat TH promoter and a
modified jellyfish (Aequorea victoria) green fluorescent protein gene is integrated. The GFP
reporter is modified by different point mutations to enhance brightness and reduce toxicity
Phenotype: No obvious changes in phenotype.
Detection of genetically modified animals: Primarily by genomic PCR using primers which
detect the EGFP gene.
References: Produced in the Allen laboratory, The Babraham Institute, Cambridge, UK. Not
yet published.
Other names: Mus musculus (animal, vertebrate, mammal, mouse, GM, mice)
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8.
Mus musculus (Linnaeus 1758) strains C57Bl6/J and C57Bl6/J x CBA/Ca,
Rosa26-Cre reporter (GMC02006)
Organism description:
Laboratory mice with an expression cassette containing the lacZ gene preceded by a lox-Pflanked “stop” sequence integrated into the host mouse ROSA26 locus.
Modification introduced: An expression cassette that contains the lacZ gene preceded by a
lox-P-flanked stop sequence was integrated into the mouse ROSA26 locus. When this strain is
crossed with a Cre recombinase-expressing mouse strain, the offspring express lacZ in the
cells in which Cre recombinase is active.
Phenotype: The mice exhibit no obvious changes in phenotype.
Detection of genetically modified animals: Primarily by genomic PCR using primers
specific to the incorporated expression cassette.
References: Soriano P. (1999) Generalized lacZ expression with the ROSA26 Cre reporter
strain. Nature Genetics 21, 70-71.
Other names: Mus musculus (animal, vertebrate, mammal, mouse, GM, mice)
9.
Mus musculus (Linnaeus 1758) strains C57Bl6/J and C57Bl6/J x CBA/Ca,
GABA-A receptor gamma2 subunit-Flox (GMC02006)
Organism description:
Laboratory mice with an integrated DNA sequence containing the loxP consensus sequences
(Flox) introduced into the mouse GABAA receptor gamma 2 subunit gene.
Modification introduced: The loxP consensus sequence, recognised by Cre recombinase, is
introduced into exons encoding the GABA-A receptor gamma 2 subunit gene.
Phenotype: No obvious changes in phenotype.
Detection of genetically modified animals: Primarily by genomic PCR using primers
specific to the integrated sequences.
References: Produced in the Luscher laboratory, Pennsylvania State University, USA. Not
yet published.
Other names: Mus musculus (animal, vertebrate, mammal, mouse, GM, mice)
10. Mus musculus (Linnaeus 1758) strains C57Bl6/J and C57Bl6/J x CBA/Ca,
KCC2-Flox (GMC02006)
Organism description:
Laboratory mice with an integrated DNA sequence containing the loxP consensus sequences
(Flox) introduced into the mouse potassium-chloride co-transporter 2 (KCC2) gene.
Modification introduced: LoxP consensus sequences, recognised by Cre recombinase, are
introduced to flank exons encoding the KCC2 gene.
Phenotype: No obvious changes in phenotype.
Detection of genetically modified animals: Primarily by genomic PCR using primers
specific to the integrated sequences.
References: Produced in the Jentsch laboratory, Hamburg University, Germany. Not yet
published.
Other names: Mus musculus (animal, vertebrate, mammal, mouse, GM, mice)
Environmental Risk Management Authority Decision: Application GMC02006
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11. Mus musculus (Linnaeus 1758) strains C57Bl6/J and C57Bl6/J x CBA/Ca,
CREB-Flox (GMC02006)
Organism description:
Laboratory mice with an integrated DNA sequence containing the loxP consensus sequences
(Flox) introduced into the mouse cAMP-response element binding protein (CREB) gene.
Modification introduced: LoxP consensus sequences, recognised by Cre recombinase, are
introduced to flank exons encoding the CREB gene.
Phenotype: No obvious changes in phenotype.
Detection of genetically modified animals: Primarily by genomic PCR using primers
specific to the integrated sequences.
References: Casanova E, Fehsenfeld S, Mantamadiotis, Lemberger T, Greiner E, Stewart Af,
Schutz G. (2001) A CamKIIalpha iCre BAC allows brain specific gene inactivation. Genesis
31, 37-42.
Other names: Mus musculus (animal, vertebrate, mammal, mouse, GM, mice)
12. Mus musculus (Linnaeus 1758) strain C57Bl6/J, ER-alpha-KO (GMC02006)
Organism description:
Laboratory mice with insertion of a neomycin resistance gene into exon 2 of the mouse
estrogen receptor alpha gene (with a functional deletion of the estrogen receptor (ER) alpha
gene).
Modification introduced: The neomycin resistance gene was inserted into exon 2 of ER
alpha resulting in the complete absence of normal ER alpha transcripts in homozygous mice.
Phenotype: Female mice homozygous for the ER alpha gene mutation are infertile as a result
of dysfunction in their gonadal axis. Male mice are sub-fertile. Heterozygous males and
females exhibit normal fertility.
Detection of genetically modified animals: Genomic PCR is used to detect the presence of
the neomycin gene, generating a 550bp product not present in the wild-type mouse.
References: Lubahn DB, Moyer JS, Golding TS, Couse JF, Korach KS, Smithies O. (1993)
Alteration of reproductive function but not prenatal sexual development after insertional
disruption of the mouse estrogen receptor. Proc. Nat. Acad. Sci. 90, 11162-11166.
Other names: Mus musculus (animal, vertebrate, mammal, mouse, GM, K/O, mice)
13. Mus musculus (Linnaeus 1758) strain C57Bl6/J, ER-beta-KO (GMC02006)
Organism description:
Laboratory mice with insertion of a neomycin resistance gene into exon 3 of the mouse
estrogen receptor beta gene (with a functional deletion of the estrogen receptor (ER) beta
gene).
Modification introduced: The neomycin resistance gene was inserted into exon 3 of ER beta
resulting in the complete absence of normal ER beta transcripts in homozygous mice.
Phenotype: Female mice homozygous for the ER beta gene mutation are sub-infertile as a
result of dysfunction in their gonadal axis. Male homozygous mice and heterozygous mice of
both sexes exhibit normal fertility.
Detection of genetically modified animals: Genomic PCR is used to detect the presence of
the neomycin gene. A forward primer in exon 3 and reverse primer in the neomycin gene
generate a 400bp product not present in the wild-type mouse.
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References: Krege JH, Hodgin JB, Couse JF, Enmark E, Warner M, Mahler JF, Sar M,
Korach KS, Gustafsson JA, Smithies O. (1998) Generation and reproductive phenotypes of
mice lacking estrogen receptor beta. Proc. Nat. Acad. Sci. 95, 15677-15682.
Other names: Mus musculus (animal, vertebrate, mammal, mouse, GM, K/O, mice)
Environmental Risk Management Authority Decision: Application GMC02006
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Annex 3: Qualitative scales for describing adverse &
beneficial effects
The following qualitative scale has been used to describe the likelihood of an adverse or
beneficial effect occurring:
Table 1:
Likelihood of effect
Descriptor
Description
Very unlikely
Not impossible, but only occurring in exceptional
circumstances
Unlikely
Could occur, but is not expected to occur under normal
conditions
Equally likely or unlikely
50:50 chance of occurring
Likely
Will probably occur at some time
Very likely (almost certain) Is expected to occur
The following qualitative scale has been used to describe the magnitude (or measure of the
severity) of an adverse effect occurring:
Table 2:
Magnitude of adverse effect
Descriptor
Examples of descriptors for type and extent of adverse effect
Minimal
Slight or insignificant, repairable or reversible, very localised (affecting
only a few individuals, single plants/animals or individual businesses), no
flow-on effects, acute rather than chronic, not affecting native or valued
species
Minor
Small, reversible and short term, localised to small land area or local
community, acute, possible affecting valued species but not native species
Moderate
Medium or mid range, largely but not completely reversible or medium
term effect, some limited flow-on effects, slight effect on native species,
affecting plants/animals/people/small industry over a wide area, but not
necessarily over the whole country
Major
Large, long term effect, but no species loss, affecting the whole country,
both acute and chronic health effects possibly leading to small number of
deaths or reduced life expectancy
Massive
Huge and widespread, irreversible, national impact, considerable secondary
effects, acute and chronic health effects leading to deaths, species loss,
serious social and cultural damage with displacement of persons and loss of
livelihood, major economic disaster
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The word scale used to describe the likelihood of beneficial effects is the same as stated in
Table 1 above. The following qualitative scale has been used to describe the magnitude (or
expected value) of a beneficial effect occurring:
Table 3:
Magnitude of beneficial effect
Descriptor
Examples of descriptors for type and extent of beneficial effect
Minimal
slight or insignificant, short term, very localised (affecting only a few
individuals, single plants/animals), no flow-on effects
Minor
small, reversible, localised to small land area, a group of individuals,
a single company/organisation or a local community
Moderate
medium or mid range, medium term, affecting
plants/animals/people/small industry over a wide area, but not
necessarily over the whole country, some flow-on effects, regional
short/medium term reduction in a weed/pest
Major
large, affecting large communities and industries, some national
impact
Massive
huge and widespread, long term, national impact, extensive
secondary or flow-on effects, eradication of a weed/pest, large
increases in employment, development of a new industry
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