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GMM Risk
Assessment
Guidance
Doc No: 1
Issue: Final
Date: November 2009
Page: 1 of 8
Guidance to completing the Genetic Modification of Micro-organisms
(GMM) Risk Assessment
…No person shall undertake any activity involving genetic modification of micro-organisms
unless… a suitable and sufficient assessment of the risks… to human health and the
environment has been carried out (Regulation 6 of the Genetically Modified Organisms
(Contained Use) Regulations 2000.
The Genetically Modified Organisms (Contained Use) Regulations 2000 and the
Environmental Protection Act 1990 (EPA) require that suitable and sufficient
assessment of the risks to human health and the environment be carried out for
activities involving the genetic modification of organisms. The primary role of the risk
assessment is to determine the appropriate control measures that are needed to
afford maximum protection to both human health and the environment. This, in turn,
will determine the notification requirements for the proposed work.
This guidance document is based on the Scientific Advisory Committee on Genetic
Modification
(SACGM).
Full
SACGM
guidance
is
available
via
http://www.hse.gov.uk/biosafety/gmo/acgm/acgmcomp/, in particular Part 2 for risk
assessment to human and animal health and Part 4 for plant health.
The final risk assessment must contain enough background and detail to ensure that
a reviewer with limited understanding of the precise nature of the work will not
require further information to comprehend the nature of any hazards. All feasible
potential hazards should be acknowledged and information should be based on
established scientific knowledge where available, duly referenced. Any uncertainty
should be acknowledged and dealt with appropriately – lack of evidence does not
equal lack of hazard.
It is a legal requirement for risk assessments to be kept for 10 years after the work
has ceased. Storage of materials is classed as active work.
For a glossary of abbreviations used in the form please go to the end of this
guidance.
Guidance on specific questions
Short description of project aim, rationale, work procedures
Outline and justify context of the research and main procedures, especially where
unusual methods are employed. If hazardous (for definition of ‘hazardous’ in this
context see SACGM guidance) biological agents are used please give reason for not
using less hazardous agents. NB: A comprehensive list of all hazardous agents is not
required in this box.
Details of insert(s) and host/vector system(s):
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Provide details for each insert, including short functional description and originating
organism/strain. In which form are these inserts received on site (e.g. live culture;
frozen tissue; naked DNA complete genome; naked DNA PCR product).
An adequate overview of the different types of GMM that will be constructed must be
provided. Host/vector systems must be given in sensible groupings with only those
combinations given which will actually be produced during the work.
Provide details of host strains with relevant disabling mutations. These can be found
in Maniatis (J. Sambrook, E. F. Fritsch and T. Maniatis (1989) Molecular Cloning, A
Laboratory Manual) or the supplier information data sheets. Where strains are
derived from a common originating strain only the latter needs to be detailed. For
example ‘E. coli BL21 and derivatives’ will include strains like Rosetta.
Details of vectors should be provided in a similar way. For example, ‘pBR322 and
derivatives’ will include P426 and pPIC 9k.
Derivatives which have disablements restored or have added functionality which may
convey a hazard must be detailed specifically.
Where a project includes multiple hosts for genetic material, e.g. cloning of genes
into bacteria and then into mammalian cells for further studies, all individual GMMs to
be created must be listed.
If relevant, a diagrammatic representation of vectors/inserts can be attached to this
form.
Overall nature of intended GMM
SACGM recommends you complete the assessment for the most at-risk group (either
human health or environment) first.
Guidance on Part A – Risk Assessment for Human Health
The objective is to identify all plausible hazards to human health and then to assess
the likelihood and potential severity of the consequences, should the hazards be
realised.
Answers should never be a stand-along ‘yes’ or ‘no’. Always give reasoning and
explanations and where possible back this up with scientific evidence and references.
A1. Mechanisms by which the GMM might pose a hazard to health
This section involves identifying those features of the GMM that have the potential to
cause harm and the mechanisms by which these hazards could be realised.
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Factors to be considered include pathogenicity of the recipient microorganism, the
properties of the sequences to be inserted and the final GMM. Areas of uncertainty
must be highlighted.

What are the hazards associated with the recipient strain(s)?
For example, for pathogens give a short description of disease symptoms,
transmission route, available prophylaxis etc.
Particular care must be given to the assessment of GMMs that have the potential to
enter human cells or establish an infection in human hosts.
Are any disabling mutations likely to be complemented (e.g. by insert/vector
combinations or recombination with wild type) or reversed by mutation? A
consequence of a reversion event in an attenuated or disabled recipient could be the
generation of a pathogenic strain that expresses the inserted gene.

What hazards are associated with the inserted genetic material?
For example from which organism / strain does it originate; is the insert potentially
harmful and in what form can it cause harm (e.g. toxins, cytokines, growth factors,
allergens, hormones or oncogenes – consider kinetics and expected level of
expression; consequences of exposure). Inserted genes can also encode
pathogenicity or virulence in other ways (invasion, surface determinants such as pili,
LPS and capsule affecting infectivity and virulence of the bacterial host).

Have the pathogenic traits of the recipient strain been altered?
Many modifications will not involve genes with products that have activities that will
be directly harmful, but adverse effects may nevertheless arise as the result of
exacerbation or alteration of existing pathogenic traits.
E.g. describe modifications; altered virulence; altered tissue tropism or host range;
altered response to prophylaxis; any modifications affecting the infectivity or
virulence of the host e.g. modified regulatory components may have unforeseen
ramifications; any modifications altering susceptibility to the immune system (e.g.
expressing interleukin 4 may inhibit the appropriate immune response to certain
pathogens).

Could the GMM or other organisms acquire harmful sequences?
There are many mechanisms by which sequences may be transferred between microorganisms and the factors that affect the frequency of such events and the likelihood
of a harmful consequence are complex. Such issues must be carefully considered in
the risk assessment. During the hazard identification process, it is important to
consider the potentially harmful consequences of sequences inserted into a GMM
being transferred to other organisms, or that the GMM itself may acquire sequences
that increase its pathogenicity.
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‘Harmful’ in this sense is anything that is either inherently harmful (e.g. toxin) or
could render a pathogen more harmful or with a selective advantage (e.g. if a
modified insert were to recombine back into a wild type pathogen. Would the inserted
sequence give an advantage to naturally occurring pathogens (e.g. drug-resistance
markers give a selective advantage and are classed as ‘harmful sequences’ if
mobilised to pathogens – therefore non-mobilisable plasmids should be used as a
general rule)?
The longer the microorganism can survive outside lab conditions the more chance
there is for a transfer event (e.g. some E coli K12 can survive for up to several days
in the gut and for similar lengths of time in the environment). Transformation and
conjugation involving plasmid DNA could result in transfer of harmful sequences
between bacteria.
If the wild type strain of the GMM is handled in the same lab this should be taken into
consideration. It is usually safest to insert sequences close to a disabling mutation so
that if recombination were to occur both would be lost at the same time – justify if a
different insertion site is chosen.
A.2 Likelihood that the GMM will be a risk to human health and safety
In this section the likelihood of the (theoretical) hazards identified in section A.1
being manifested should be realised, assessed and their potential impact understood.

Likelihood: Could the GMM establish an infection in vivo and how
efficient would it be?
Consider the overall fitness of the GMM (ability to establish an infection, how efficient
that infection would be and its ability to spread within a host or through a
community) and the probability that rare events like mutation, recombination or
gene transfer may occur (published data of numerical frequencies may exist, e.g.
comparing the frequency of transfer of mobilisable, mobilisation defective and nontransferable plasmids). Is there selective pressure for deletion of the modified
sequences or is there a vital sequence nearby preventing this?

How severe might the consequences be?
For example consider the severity of harm to an individual and the severity of
secondary spread to the wider community; consider availability of prophylaxis.
Consider if there are any particular groups of individuals who might be at higher risk
than others and whether they are likely to come into contact with the work.

Containment level needed to sufficiently protect human health
At this stage the setting of a minimal containment level to protect human health is
only provisional and estimated to protect those who come into contact with the GMM.
Consider the containment level necessary to control the risk of the recipient microorganism and make a judgement about whether the proposed modification will result
in a GMM that is more hazardous, less hazardous or approximately equivalent.
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Guidance on Part B – Risk Assessment for the Environment
There is a requirement under both the Genetically Modified Organisms (Contained
Use) Regulations 2000 and the Environmental Protection Act 1990 to consider risks
to the environment. The objective of this section is to identify all plausible hazards to
the environment (animal, plant and microbial) and to determine the likelihood and
the possible consequences of an accidental release of a GMM from containment into
the environment (including waste disposal, equipment failure and human spread).
Consider the local as well as the wider environment.
GMMs with the potential to infect or colonise animals and plants are of primary
concern.
If a GMM is incapable of infecting or impacting on any species other than humans this
should be stated here with supporting evidence.
B1. Mechanisms by which the GMM might pose a hazard to the environment
This section focuses on the identification of those features of the GMM that have the
potential to cause harm to the environment and the mechanisms by which theses
hazards could be realised.

Hazards associated with the recipient strain(s)
Consider pathogenicity, infectivity, toxicity, virulence, allergenicity, colonisation,
parasitism, symbiosis and competition. How does the proposed modification affect
these traits? Take account of the DEFRA (SAPO) classification of animal pathogens:
www.defra.gov.uk/animalh/diseases/pathogens/classification.htm. GMMs based on
such pathogens may require DEFRA licenses to handle (under SAPO) or import
(under IAPO). Also consider the DEFRA Plant Health Division for plant pathogens:
http://www.defra.gov.uk/planth/ph.htm .

Has the stability or survivability of the recipient been altered?
Consider modifications compared to wild type; longevity inside / outside the host. If
the GMM is short-lived this normally provides safe ‘biological containment’. Consider
viability within organisms as well as in soil, water or other sites, vegetative as well
replicating stage.

Hazards associated with genetic inserts – Does the inserted gene pose
a risk to other organisms?
GMMs might be a hazard to the environment by virtue of the properties inherent to
the genetic insert, even if the recipient micro-organism poses no specific risk, for
instance pesticidal genes.
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Hazards arising from the alteration of traits in the GMM
For example, consider alteration of stability or survivability (enhanced resistance to
oxidative stress, UV, temp fluctuations or dehydration); pathogenicity or infectivity
(altered susceptibility to host immune systems); tissue tropism or host range.

Could the GMM or other organisms in the environment acquire harmful
sequences?
Consider exchange of genetic material with other strains/pathogens in both
directions, from GMM to environment but also from environment to GMM, making it
more harmful e.g. E coli K12 during persistence in gut, especially under bacterial
stress conditions).
Can any disabling/attenuating mutations be reverted or complemented? In particular,
assess the possibility of sequence mobilisation in bacteria (e.g. through [re]mobilisable plasmid vectors).
B2. Likelihood that the GMM will be a risk to the environment
In this section the chances of the hazards identified in section B1 being realised
should be evaluated and understood.

What is the likelihood that the hazard(s) will be manifested?
Where the likelihood of harm is poorly understood, a precautionary approach should
be adopted until evidence to the contrary has been obtained.
Estimate the risk as high, medium, low or negligible, with justification.

What will be the consequences if the hazard is realised?
Even if a quantitative estimate is not possible, a qualitative appraisal of the impact
on other species or ecosystems should be possible. Consequences could be described
as severe, modest, minor or negligible, with justification. Please note that even if the
likelihood of occurrence is low the potential consequences may be severe.

Containment level needed to protect the environment
This requires a judgement as to whether the measures required for the recipient
strain in the appropriate table of containment measures in Schedule 8 of the GMOs
(Contained Use) Regulations 2000 are also appropriate for the GMM. If some
measures are not needed or extra measures are required the containment level
should be adjusted accordingly. Requiring a measure listed as a higher containment
level will automatically upgrade the final containment level for the work.
Guidance on Part C. Review of Procedures and Control Measures
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The requirements of the final containment level must be sufficient to control al the
potential harmful properties of the GMM and offer sufficient protection for both
human health and the environment. Any non-standard operations increasing the risk
of exposure or likelihood of release must be taken into account to decide whether
additional containment and control measures are required.
Implement measures to safeguard human health and the environment

What operations and procedures might increase risk of exposure?
Consider for example if airborne transmission arising from aerosol generating
procedures will pose risk of exposure; use of sharps.

What control measures and monitoring procedures are to be used in
addition to GM (Contained Use) Regulations Schedule 8?
Consider (in outline) the use of Microbiological Safety Cabinets; centrifugation inside
removable sealed rotors; fumigation; etc. Full procedures should be appended to the
risk assessment including training undertaken.

Are the potential
managed?
routes
of
environmental
release
known
and
Consider for example aerosols and contaminated waste; details of disinfection
procedures including validated effectiveness (log reduction) and waste disposal.
Guidance to Part D. Activity Class
Assign final GM activity class – 1, 2, 3 or 4
The final GM activity class should be suggested by the applicant and will be subject to
GMSC assessment.
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Date: November 2009
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Glossary of Abbreviations
BSC
Biological Safety Committee (in this case equivalent to the GMSC)
BSO
Biological Safety Officer
DEFRA
Department of Environment, Food and Rural Affairs
EPA
Environmental Protection Act 1990
GM
Genetic Modification
GMM
Genetically Modified Micro-organism
GMO
Genetically Modified Organism
GMSC
Genetic Modification Safety Committee
HSE
Health & Safety Executive
SACGM
Scientific Advisory Committee on Genetic Modification
SAPO
Specific Animal Pathogens Order 1998
IAPO
Importation of Animal Pathogens Order 1980
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