Download Amendments - Office of the Vice Provost

FOR IBC OFFICE USE ONLY
AMENDMENT #
APPROVAL DATE
Tufts University & Tufts Medical Center
Institutional Biosafety Committee (IBC)
Tel: 617-636-4109, Fax: 617-636-8354
E-mail: [email protected]
Website: http://viceprovost.tufts.edu/ibc/
To submit for review, please e-mail this form to the IBC Office ([email protected])
AMENDMENT FORM
Please note that amendments may also affect other aspects of that registration, and those areas should also be reflected
in this document. Some amendments may require submission to the full Institutional Biosafety Committee. Others may
be approved administratively by a Biosafety Officer. The Biosafety Officers reserves the right to call to full Committee
review of the amendment or to require a new registration instead, if necessary. When submitting an amendment, the
Principal Investigator is required to review all of the details of the original registration to assure the IBC that all
unamended details remain identical to the original registration. Complete this form and send it electronically to [email protected] to submit for review.
Amendment to Registration #
Registration Title
Mark the checkboxes as confirmation:
I am familiar with and agree to abide by the NIH Guidelines for Recombinant and Synthetic Nucleic Acid
Molecules, CDC/NIH Biosafety Guidelines, OSHA Standards, institutional policies, and other federal, state
and local regulations relating to this project.
I attest that the information contained in the attached registration is accurate and complete.
I accept responsibility for ensuring that all personnel involved in this project will be trained regarding the
procedures approved, the potential biohazards, relevant biosafety practices, and emergency procedures. I
confirm that the relevant Exposure Response Plan(s) will be followed.
I will submit written reports to the Institutional Biosafety Committee concerning:
1. Any accident that results in a known or potential exposure to recombinant or synthetic nucleic acid
materials, infectious agents or biological toxins; or any incident resulting in the known or suspected
release into the environment of recombinant or synthetic nucleic acid materials, infectious agents or
biological toxins into the environment.
2. Any problems with physical or biological containment safety procedures or equipment, or facility
failures.
3. Any new information bearing on the safety of this work such as technical data relating to hazard and
safety procedures.
Electronic Signature of
the Principal Investigator:
Date:
By typing your name you are submitting an electronic signature that confirms your understanding and adherence to the
above statements and IBC policies. This is considered legal documentation and confirmation of your agreement to execute
all activities as approved.
1
INSTRUCTIONS FOR COMPLETING THIS FORM
- Please type responses within the space/box provided. You can mark the checkboxes by double-clicking on the box.
- Submit the completed draft electronically to the IBC Office e-mail ([email protected])
- A Biosafety Officer will pre-review the form and determine if the research requires IBC Full Committee Approval or IBC
Administrative Approval. Please be aware that it is in your best interest to submit the draft to the IBC office well in
advance of the submission deadline for the IBC meeting.
I. PROPOSED MODIFICATION(S)
More than one of the Sections below may be affected by the proposed modification. Please complete each section as
necessary.
Addition/modification of where agent(s) is/are used (any new space must be inspected prior to use)
Addition/modification in use of rDNA or synthetic DNA
Addition of biological materials (infectious agents, biological toxins, select agents/toxins, and human material)
Addition/modification of procedures with live animals
Nontechnical Summary of Proposed Modification(s)
II. ADDITION/MODIFICATION OF LOCATION
List ONLY NEW Laboratories/Facilities where research is to be conducted and the corresponding biosafety level. Include
cold/warm rooms, equipment rooms, and location(s) of biosafety cabinets (BSC). All new spaces must be inspected by a
Biosafety Officer prior to use. For areas within the centralized animal facilities where the work will be performed, please
indicate “DLAM” (Department of Laboratory Animal Medicine), “LAMS” (Laboratory Animal Medicine Services),” or
“CBU” (Comparative Biology Unit), as appropriate. Provide housing levels in the section below.
Room Purpose
Building and Room Number for Labs
Biosafety Level
(Main Lab, Storage, Tissue Culture,
Procedure, etc.)
BSL-1
BSL-1
BSL-1
BSL-1
BSL-2
BSL-2
BSL-2
BSL-2
BSL-3
BSL-3
BSL-3
BSL-3
N/A
N/A
N/A
N/A
Below list ALL locations where work with live animals will be housed. ABSL refers to containment of the animal. For
areas within the centralized animal facilities, please indicate “DLAM” (Department of Laboratory Animal Medicine),
“LAMS” (Laboratory Animal Medicine Services),” or “CBU” (Comparative Biology Unit), as appropriate.
Room Purpose
Building and Room Number for Labs
(Infection, Monitoring, Other
procedures, Satellite Housing, etc.)
Biosafety Level
ABSL-1
ABSL-2
ABSL-3 N/A
ABSL-1
ABSL-2
ABSL-3 N/A
ABSL-1 containment with filter-top
cages and the use of “Do Not Disturb”
cage cards 72 hours after agent
administration
2
Please Note: All locations will be inspected by a Biosafety Officer, therefore if there are any locations that should be
REMOVED from your registration, please send an email to the IBC Office. A Biosafety Officer may follow up with you for
further close out or decommissioning procedures.
III. ADDITION/MODIFICATION OF RECOMBINANT OR SYNTHETIC DNA
A. Type of Modification
Addition of recombinant or synthetic DNA (new vectors, etc.)
Change in use of recombinant or synthetic DNA
B. Justification and Technical Details
1. Describe the purpose and experimental details for the change:
2. Name(s), Relevant Biosafety Level(s), and NIH Guidelines Categorization(s) of DNA to be added
Name
Section(s) of NIH Guidelines
Biosafety Level
BSL-1
ABSL-1
BSL-1
ABSL-1
BSL-1
ABSL-1
BSL-2
ABSL-2
BSL-2
ABSL-2
BSL-2
ABSL-2
BSL-3
ABSL-3
BSL-3
ABSL-3
BSL-3
ABSL-3
3. Nature and species of sequence(s) added
Name of Sequence
Source
(species name)
Nature of Insert or
Expressed Protein
Function
Promoter
4. New vectors and host cells
Name of Vector
Gene Transfer
Method
Source of Vector
Name of Host for
Propagation or of
Packaging Cell Lines
Target Recipient of
rDNA
5. Provide additional information for sequences that code for toxins, oncogenes, increased virulence, or other
potentially hazardous RNA/protein.
6. Will any of the constructs contain more than 50% of the genome for a virus (or viral family)? If so, please provide
more detail below.
7. Provide restriction maps of vectors unless they are commercially available. If commercially available, please indicate
vendor.
9. For any viral vector in use, indicate whether it generates a replication competent product and indicate the assay
system used to measure virus titer. Please refer to the “Policy on Retroviral Replication Competency Testing.”
NOTE: Insertion of high risk genes (e.g. oncogenes) also requires testing (please describe below).
10. Please confirm that results of testing will be available for review.
11. Is a helper virus required for replication?
NO
YES
NO
YES
3
12. Describe the assay used to measure titer of replication competent virus (background) generated or helper virus.
13. What is the percent of synthetic DNA in the construct and anticipated function?
C. USE OR ACQUISITION OF GENETICALLY MODIFIED ORGANISMS
 This section is necessary for both use of - and any acquisition of (transfer, purchase, etc.) - genetically modified
invertebrates, vertebrates, or plants.
 Any creation of genetically engineered organisms must be described in the Experimental Details section.
 Exempted from this section is the acquisition, use, or breeding of genetically engineered rodents at ABSL-1. That
information should be provided in the IACUC form only.
See “IBC Policy on Genetically Engineered Mutants” for detailed information about the IBC review of genetic mutants and
which type of review is required.
1. Please describe how recombinant or synthetic nucleic acid molecules will be used.
2. Please check the applicable box and complete the sections below for insects or plants.
Use/generation of genetically engineered INSECTS ONLY
Use/generation of genetically engineered PLANTS ONLY
3. List the insect or plant species to be used:
4. Transgene(s) Information
Transgene Name Or
Family
Transgene
Function
Transgene Source
Vector Used
Recipient Strain
5. What is the method of transformation?
6. Does the gene encode a toxin or other hazardous agent? If yes, please describe.
7. Transgenic agents cannot be released into the environment. Please describe methods to prevent release (e.g. fly trap)
and methods of disposal:
IV. ADDITION OF BIOLOGICAL MATERIALS
A. ADDITION OF INFECTIOUS AGENT(S) – Altered viruses, bacteria, and cell lines should be reserved for
Section III. Human source materials (e.g. blood or cell lines) should be reserved for Section IV Part D.
1. Describe the purpose and experimental details for the infectious agent addition:
2. Name(s), Relevant Biosafety Level(s), and Pathology Information
Pathology of Agent
Name
Biosafety Level
BSL-1
BSL-1
BSL-1
BSL-2
BSL-2
BSL-2
Human pathogen, Animal pathogen,
Both, Neither or N/A
BSL-3
BSL-3
BSL-3
4
SELEC
3. If a human pathogen, what is the infectious dose for a healthy human adult?
4. Is the agent attenuated or replication deficient?
NO
YES
5. If yes, please describe in detail the testing that will be done to confirm attenuation or replication incompetence.
Include the location of records.
6. Please confirm that results of testing will be available for review.
YES
NO
B. ADDITION OF BIOLOGICAL TOXIN(S)
1. Describe the purpose and experimental details for the biological toxin addition:
2. Name(s), Relevant Biosafety Level(s), and Toxicity Information
Toxicity of Agent
Name
Biosafety Level
BSL-1
BSL-1
BSL-1
BSL-2
BSL-2
BSL-2
To humans, To animals, To both,
Neither or N/A
BSL-3
BSL-3
BSL-3
3. What is the maximum quantity of toxin that will be present?
4. What is the LD50 of the toxin in humans?
5. What is the LD50 of the toxin in animal species used in this experiment?
6. How will the laboratory track inventory of the toxin?
7. How will the laboratory inactivate the toxin?
C. ADDITION OF SELECT AGENT(S) OR TOXIN(S) - To determine if your study falls within the Select Agent Rule,
refer to http://www.selectagents.gov/SelectAgentsandToxins.html
1. Describe the purpose and experimental details for the select agent and/or toxin addition:
2. Name(s), Relevant Biosafety Level(s), and Toxicity or Pathology Information
Name
Pathology or Toxicity
Biosafety Level
BSL-1
BSL-2
Humans, Animals, Both, Neither, or N/A
BSL-3
3. If a human pathogen, what is the infectious dose for a healthy human adult?
4. If a toxin, what is the maximum quantity that will be present?
5. What is the LD50 of the toxin in humans?
6. What is the LD50 of the toxin in animal species used in this experiment?
D. ADDITION OF HUMAN SOURCE MATERIAL (HSM)s/ NON-HUMAN PRIMATE SOURCE MATERIAL – Please
review the IBC website for guidance on what HSMs to include in this form.
1. Please check all that apply below and complete the relevant details in the boxes provided.
I will use human source materials in animals.
I will use human/non-human primate (NHP) cells as recipient host cells.
5
I will use modified human cells, but I will not make the modifications to these cells.
Other
2. Describe the purpose and experimental details for the human source material(s) addition:
HUMAN/NON-HUMAN PRIMATE (NHP) BLOOD, BODY FLUIDS, TISSUE, ORGANS
Name
Biosafety Level
BSL-1
BSL-1
BSL-1
HUMAN/NON-HUMAN PRIMATE (NHP) CELL LINES
Name
BSL-2
BSL-2
BSL-2
BSL-3
BSL-3
BSL-3
Biosafety Level
BSL-1
BSL-1
BSL-1
BSL-2
BSL-2
BSL-2
BSL-3
BSL-3
BSL-3
V. ADDITION/MODIFICATION OF PROCEDURES WITH LIVE ANIMALS
Complete this section ONLY if the biological material(s) listed on this amendment will be administered to animals.
Please provide a response for all items. If a question does not apply, please write “N/A.” Section can be duplicated as
necessary.
1. Please check
to confirm the information is the same as described in the original registration. If different answers
are applicable for this amendment, please complete the questions below.
A. ADMINISTRATION OF AGENT
1. Name of species to be exposed to agent:
2. Are the animals immunosuppressed?
No
Yes
3. Agent(s) and dose(s) administered:
4. Route(s) of administration:
5. Will administration(s) be done inside a biosafety cabinet?
Yes
No
6. Will the biological agent or hazardous metabolite be excreted by urine, feces or wound drainage? Provide
duration, if applicable.
7. Will a DLAM/LAMS/CBU biohazard cage card be used to identify the agent used?
If no, will an alternative method be used to identify the biohazardous agent?
Yes
No
No
Yes
Describe alternate method, if applicable
B. TRANSPORTATION OF ANIMALS
Will infected live animals be transferred to/located in areas outside the DLAM/LAMS/CBU facilities?
Yes
No
If yes, please describe here how transportation will be handled in order to minimize biohazardous risk. Please
include the location in Section II.
C. DECONTAMINATION OF ANIMAL USE
Will handling and disposal of bedding, cages, and animal carcasses be done by DLAM/LAMS/CBU staff in
6
accordance with standard ABSL-appropriate DLAM/LAMS/CBU procedures?
Yes
No
a. If no, will handling and disposal of bedding, cage, and animal carcasses be done by laboratory staff?
No
Yes
b. Describe lab staff’s method, if applicable.
VI. RISK ASSESSMENT
1. Please check
to confirm the information is the same as described in the original registration. If different answers
are applicable for this amendment, please complete the questions below.
A. Hazardous processes used with agent. Check all that apply.
Centrifuge
Animal Model
Tissue harvesting
Tissue homogenization
Sharps: If yes, please describe:
Sonication
Pipetting
Cell sorting (flow cytometry or other method)
Other: If yes, please specify:
B. Exposure route of agents. Check all that apply.
Ingestion
Percutaneous
Other: If yes, please specify:
Mucous Membrane
Inhalation
C. The risk of exposure will be mitigated by the following:
Check all that apply and indicate with what agent each is used and the location of use (lab room #, animal facility, etc.).
If using engineering controls, provide the date of certification.
Personal Protective Equipment
Gloves
Lab Coat
Disposable Lab Gown
Disposable Booties
Tyvek Suit
N-95 Respirator
Surgical Mask
Powered Air Purifying Respirator (PAPR)
Safety Glasses
Other: (specify)
Engineering Controls
Biosafety Cabinet (BSC)
Other: (specify)
Agent(s) and Location of Use
Date of Certification
VII. DECONTAMINATION
If the amendment contains more than one agent, provide clarification for each of the answers below.
1. Please check
to confirm the information is the same as described in the original registration. If different answers
are applicable for this amendment, please complete the questions below.
A. What chemical and/or thermal processes will be used for decontamination?
7
B. Describe the processes involved in the decontamination and/or disposal of the following:
1. Liquid waste:
2. Contaminated solid waste:
3. Cultures, plates, stocks, etc.:
4. Biosafety cabinet and/or workspace:
VIII. EXPOSURE RESPONSE PLAN(S)
1. Please check
to confirm that the necessary Exposure Response Plan(s) is/are the same as described in the original
registration. If different plans apply to this amendment, please complete the questions below.
2. Please list the titles of ALL Exposure Response Plan(s) associated with this AMENDMENT ONLY. Many plans are
available for download on the IBC website.
A. Title of each Exposure Response Plan to be utilized:
B. OR if a specific plan has not been developed for the agent you are proposing to work with or if you require changes to
the existing Exposure Response Plan, the Biosafety Officer will work directly with you at the time of pre-review to
develop a specific Plan, if necessary. To aid in this process, please provide the following information:
1. Are there signs and symptoms of exposure?
2. Identify prophylactic medicines or vaccinations for this agent, if available.
3. Is there an increased risk to immunocompromised individuals exposed to this agent?
IX. DUAL USE RESEARCH OF CONCERN (DURC)
Life sciences research that, based on current understanding, can be reasonably anticipated to provide knowledge,
information, products, or technologies that could be directly misapplied to pose a significant threat with broad potential
consequences to public health and safety, agricultural crops and other plants, animals, the environment, material, or
national security.
1. Agent or Toxin Involved in Project (Check All That Apply)
Please verify if this project directly involves nonattenuated forms of 1 or more of the 15 listed agents. If this project
does not involve any of the listed forms below, please indicate the “Not Applicable” box:
Bacteria:
Viruses:
8
Bacillus anthracis
Burkholderia mallei
Burkholderia pseudomallei
Clostridium botulinum (toxin-producing strains)
Francisella tularensis
Yersinia pestis
Toxin:
Botulinum neurotoxin (any quantity)
Avian influenza virus (highly pathogenic)
Ebola virus
Foot-and-mouth disease virus
Marburg virus
Reconstructed 1918 influenza virus
Rinderpest virus
Variola major virus
Variola minor virus
Not applicable, the proposed research does not include any of the agents listed above.
2. If you selected any of the agents listed above, please check all categories that apply to the proposed work in this
registration. See the “US Government Policy for Oversight of DURC” and NIH/OBA Educational Materials.
Increase virulence
Overcomes immunity
Develop resistance to drugs
Increase transmission
Change tropism
Increase host susceptibility
Regenerate extinct pathogens
Enhances the harmful consequences of the agent or toxin
Disrupts immunity or the effectiveness of an immunization against the
agent or toxin without clinical or agricultural justification
Confers to the agent or toxin resistance to clinically or agriculturally useful
prophylactic or therapeutic interventions against that agent or toxin or
facilitates their ability to evade detection methodologies
Increases the stability, transmissibility, or the ability to disseminate the
agent or toxin
Alters the host range or tropism of the agent or toxin
Enhances the susceptibility of a host population to the agent or toxin
Generates or reconstitutes an eradicated or extinct agent or toxin listed in
the “US Government Policy for Oversight of DURC,” Section III.2
None of the above applies
3. If you selected any of the experimental procedures listed above, please explain why this research cannot be
conducted with agents other than those listed in question #1:
OTHER RELEVANT LINKS
OVPR IBC Website:
http://viceprovost.tufts.edu/ibc/
Tufts University EHS Website:
http://publicsafety.tufts.edu/ehs/
Pathogen Safety Data Sheets:
Biosafety in Microbiological and
Biomedical Laboratories (BMBL):
NIH Recombinant DNA Guidelines:
http://www.phac-aspc.gc.ca/msds-ftss/index.html#menu
http://www.cdc.gov/biosafety/publications/bmbl5/
http://osp.od.nih.gov/sites/default/files/NIH_Guidelines.html
9
APPENDIX A - NIH Recombinant or synthetic nucleic acid molecules Categories
BIOHAZARDOUS MATERIALS REGISTRATION FORM
The NIH rDNA Guidelines identify six categories of experiments involving recombinant DNA:
(i)
those that require Institutional Biosafety Committee (IBC) approval, RAC review, and NIH Director approval
before initiation,
(ii)
those that require NIH/OBA and Institutional Biosafety Committee approval before initiation,
(iii)
those that require Institutional Biosafety Committee and Institutional Review Board approvals and RAC review
before research participant enrollment,
(iv)
those that require Institutional Biosafety Committee approval before initiation,
(v)
those that require Institutional Biosafety Committee notification simultaneous with initiation, and
(vi)
those that are exempt from the NIH Guidelines.
According to the Guidelines, the PI is responsible for identifying which category the proposed rDNA work falls under. The 6
categories are listed and defined below.
Section III-F Experiments that are exempt from NIH Guidelines but covered by local regulations. Exempt experiments must be
registered with the Tufts University IBC or the TCSVM IBC as appropriate.
III-F-1
Those synthetic nucleic acids that: (1) can neither replicate nor generate nucleic acids that can replicate in any
living cell (e.g., oligonucleotides or other synthetic nucleic acids that do not contain an origin of replication or
contain elements known to interact with either DNA or RNA polymerase), and (2) are not designed to integrate
into DNA, and (3) do not produce a toxin that is lethal for vertebrates at an LD50 of less than 100 nanograms per
kilogram body weight. If a synthetic nucleic acid is deliberately transferred into one or more human research
participants and meets the criteria of Section III-C, it is not exempt under this Section.
III-F-2
Those that are not in organisms, cells, or viruses and that have not been modified or manipulated (e.g.,
encapsulated into synthetic or natural vehicles) to render them capable of penetrating cellular membranes.
III-F-3
Those that consist solely of the exact recombinant or synthetic nucleic acid sequence from a single source that
exists contemporaneously in nature.
III-F-4
Recombinant or synthetic nucleic acid molecules that consist entirely of nucleic acids from a prokaryotic host
including its indigenous plasmids or viruses when propagated only in that host (or a closely related strain of the
same species), or when transferred to another host by well established physiological means.
III-F-5
Recombinant or synthetic nucleic acid molecules that consist entirely of nucleic acids from a eukaryotic host
including its chloroplasts, mitochondria, or plasmids (but excluding viruses) when propagated only in that host (or
closely related strain of the same species).
III-F-6
Those that consist entirely of DNA segments from different species that exchange DNA by known physiological
processes, though one or more of the segments may be a synthetic equivalent. See Appendix A-I through A-VI for a
list of natural exchangers that are exempt from the “NIH Guidelines”.
III-F-7
Those genomic DNA molecules that have acquired a transposable element, provided the transposable element
does not contain any recombinant and/or synthetic DNA.
III-F-8
Recombinant or synthetic nucleic acid molecules in experiments that do not present a significant risk to health or
the environment as determined by the NIH Director, RAC and following appropriate notice and opportunity for
public comment. See Appendix C of the NIH Guidelines.
The following classes of experiments that are exempt under Section III-F-8 Appendix C
C-1
Recombinant or synthetic nucleic acid molecules in Tissue Culture?
Recombinant or synthetic nucleic acid molecules containing less than one-half of any eukaryotic viral genome (all
viruses from a single family being considered identical – see Appendix C-VII-E, Footnotes and References of Appendix
C), that are propagated and maintained in cells in tissue culture are exempt from these NIH Guidelines with the
10
exceptions listed in Appendix C-1-A.
C-2
Escherichia coli K-12 Host-Vector Systems?
Experiments which use Escherichia coli K-12 host-vector system, with the exception of those experiments listed in
Appendix C-II-A are exempt from the NIH Guidelines provided that: (i) the Escherichia coli host does not contain
conjugation proficient plasmids or generalized transducing phages; or (ii) lambda or lambdoid of Ff bacteriophages
or non-conjugative plasmids (see Appendix C-VIII-B, Footnotes and References of Appendix C) shall be used as
vectors. However, experiments involving the insertion into Escherichia coli K-12 of DNA from prokaryotes that
exchange genetic information (see Appendix C-VIII-C, Footnotes and References of Appendix C) with Escherichia coli
may be performed with any Escherichia coli K-12 vector (e.g., conjugative plasmid). When a non-conjugative vector
is used, the Escherichia coli K-12 host may contain conjugation-proficient plasmids either autonomous or integrated,
or generalized transducing phages. For these exempt laboratory experiments, Biosafety Level 1 (BSL1) physical
containment conditions are recommended. For large scale fermentation experiments, the appropriate physical
containment conditions need not be greater than those for the host organism unmodified by techniques using
recombinant or synthetic nucleic acid molecules; the Institutional Biosafety Committee can specify higher
containment if deemed necessary.
C-III
Saccharomyces Host-Vector Systems?
Experiments involving Saccharomyces cerevisiae and Saccharomyces uvarum host-vector systems, with the
exception listed in Appendix C-III-A, are Exempt from the NIH Guidelines. For these exempt experiments, BL1
physical containment is recommended. For large scale fermentation experiments, the appropriate physical
containment conditions need be no greater than those for the unmodified host organism; the Institutional Biosafety
Committee can specify higher containment if deemed necessary.
C-IV
Kluyveromyces Host-Vector Systems
Experiments involving Kluyveromyces lactis host-vector systems, with the exception of experiments listed in
Appendix C-IV-A, are exempt from the NIH Guidelines provided laboratory-adapted strains are used (i.e. strains that
have been adapted to growth under optimal or defined laboratory conditions). For these exempt experiments, BL1
physical containment is recommended. For large-scale fermentation experiments, the appropriate physical
containment conditions need be no greater than those for the unmodified host organism; the Institutional Biosafety
Committee may specify higher containment if deemed necessary.
C-V
Bacillus subtilis or Bacillus licheniformis Host-Vector Systems?
Any asporogenic Bacillus subtilis or asporogenic Bacillus licheniformis strain which does not revert to a spore-former
with a frequency greater than 10-7 may be used for cloning DNA with the exception of those experiments listed in
Appendix C-V-A, Exceptions. For these exempt laboratory experiments, BL1 physical containment conditions are
recommended. For large scale fermentation experiments, the appropriate physical containment conditions need be
no greater than those for the unmodified host organism; the Institutional Biosafety Committee can specify higher
containment if deemed necessary.
C-VI
Appendix C-VI. Extrachromosomal Elements of Gram Positive Organisms
Recombinant or synthetic nucleic acid molecules derived entirely from extrachromosomal elements of the
organisms listed below (including shuttle vectors constructed from vectors described in Appendix C), propagated
and maintained in organisms listed below (see Guidelines for list) are exempt from these NIH Guidelines.
C-VII
The Purchase or Transfer of Transgenic Rodents?
The purchase or transfer of transgenic rodents for experiments that require BL1 containment (see Appendix G-III-M,
Footnotes and References of Appendix G) are exempt from the NIH Guidelines.
Appendix C-VIII. Generation of BL1 Transgenic Rodents via Breeding
The breeding of two different transgenic rodents or the breeding of a transgenic rodent and a non-transgenic rodent
with the intent of creating a new strain of transgenic rodent that can be housed at BL1 containment will be exempt
from the NIH Guidelines if:
(1) Both parental rodents can be housed under BL1 containment; and
(2) neither parental transgenic rodent contains the following genetic modifications: (i) incorporation of more than
one-half of the genome of an exogenous eukaryotic virus from a single family of viruses; or (ii) incorporation of a
transgene that is under the control of a gammaretroviral long terminal repeat (LTR); and
(3) the transgenic rodent that results from this breeding is not expected to contain more than one-half of an
11
exogenous viral genome from a single family of viruses.
Section III-E Experiments that require IBC notice simultaneous with initiation.
III-E
Experiments not included in Sections III-A, III-B, III-C, III-D, III-F and their subsections are considered in this section.
All such experiments may be conducted at BL 1.
III-E-1
Experiments involving the formation of recombinant or synthetic nucleic acid molecules containing no more than
2/3 of the genome of any eukaryotic virus (all viruses from a single Family being considered identical) may be
propagated and maintained in cells in tissue culture using BL 1 containment. It must be shown that the cells lack
helper virus for the specific Families of defective viruses used. The DNA may contain fragments of the genome of
viruses from more than one Family, but each fragment shall be less than 2/3 of a genome.
III-E-2
Experiments involving whole plants modified with recombinant or synthetic nucleic acid molecules, and/or
experiments involving organisms modified with recombinant or synthetic nucleic acid molecules - associated with
plants, except those that fall under Section III-A, III-B, III-C, III-D, or III-F. See Section III-E-2 for recommendation of
containment levels.
III-E-3
Experiments involving the generation of rodents in which the animal’s genome has been altered by stable
introduction of recombinant or synthetic nucleic acid molecules, or nucleic acids derived therefrom, into the germline (transgenic rodents). Only experiments that require BL1 containment are covered under this section;
experiments that require BL2, BL3, or BL4 containment are covered under Section III-D-4.
Section III-D Experiments that require IBC approval before initiation of experiments.
III-D-1
Experiments using Risk Group 2, Risk Group 3, Risk Group 4, or restricted agents as host-vector systems. (See
Section II-A Risk Assessment.)
III-D-1-a
Introduction of recombinant or synthetic nucleic acid molecules into Risk Group 2 (RG-2) agents is usually
conducted at BSL2 containment. Experiments with such agents will usually be conducted with whole animals at
BSL2 or BL2-N containment. (
III-D-1-b
Introduction of recombinant or synthetic nucleic acid molecules into Risk Group 3 (RG-3) agents is usually
conducted at BSL3 containment. Experiments with such agents will usually be conducted with whole animals at
BSL3 or ABSL-3 containment.
III-D-1-c
Introduction of recombinant or synthetic nucleic acid molecules into Risk Group 4 (RG-4) agents is usually
conducted at BSL4 containment. Experiments with such agents will usually be conducted with whole animals at
BSL4 or ABSL-4 containment.
III-D-2
Experiments in which DNA from Risk Group 2, Risk Group 3, Risk Group 4, or restricted agents is cloned into
nonpathogenic prokaryotic or lower eukaryotic host-vector systems. Experiments involving the formation of
recombinant or synthetic nucleic acid molecules for certain genes coding for molecules toxic for vertebrates
require NIH/OBA approval.
III-D-2-a
Experiments in which DNA from RG- 2 or RG- 3 agents is transferred into nonpathogenic prokaryotes or lower
eukaryotes may be conducted at BSL2 containment. Experiments in which DNA from RG-4 agents is transferred
into nonpathogenic prokaryotes or lower eukaryotes may be performed under BSL2 containment after
demonstration that only a totally and irreversibly defective fraction of the agent’s genome is present in a given
recombinant. The IBC may approve the specific lowering of containment for particular experiments to BSL1. Many
experiments in this category are exempt from the “NIH Guidelines”.
III-D-2-b
Experiments in which DNA from restricted agents is transferred into nonpathogenic prokaryotes or lower
eukaryotes shall be performed under containment conditions determined by NIH/OBA following a case-by-case
review.
III-D-3
Experiments involving the use of infectious DNA or RNA viruses or defective DNA or RNA viruses in the presence of
helper virus in tissue culture systems.
12
III-D-3-a
Experiments involving the use of infectious or defective RG-2 viruses in the presence of helper virus may be
conducted at BSL2 containment. (See Appendix B for information on Risk Groups.)
III-D-3-b
Experiments involving the use of infectious or defective RG-3 viruses in the presence of helper virus may be
conducted at BSL3 containment. (See Appendix B for information on Risk Groups.)
III-D-3-c
Experiments involving the use of infectious or defective RG-4 viruses in the presence of helper virus may be
conducted at BSL4 containment. (See Appendix B for information on Risk Groups.)
III-D-3-d
Experiments involving the use of infectious or defective restricted poxviruses in the presence of helper virus shall
be determined on a case-by-case basis following NIH/OBA review. A USDA permit is required for work with plant or
animal pathogens.
III-D-3-e
Experiments involving the use of infectious or defective viruses in the presence of helper virus which are not
covered in Sections III-D-3-a through III-D-3-d may be conducted at BSL1.
III-D-4
Experiments involving whole animals.
III-D-4-a
Recombinant or synthetic nucleic acid molecules, or DNA or RNA molecules derived therefrom, from any source
except for greater than two-thirds of eukaryotic viral genome may be transferred to any non-human vertebrate or
any invertebrate organism and propagated under conditions of physical containment comparable to BSL1 or ABSL1
and appropriate to the organism under study. Animals that contain sequences from viral vectors, which do not
lead to transmissible infection either directly or indirectly as a result of complementation or recombination in
animals, may be propagated under conditions of physical containment comparable to BSL1 or ABSL1 and
appropriate to the organism under study. Experiments involving the introduction of other sequences from
eukaryotic viral genomes into animals are covered under Section III-D-4-b. The investigator must demonstrate that
the fraction of the viral genome being utilized does not lead to productive infection.
III-D-4-b
Experiments involving recombinant or synthetic nucleic acid molecules , or DNA or RNA derived therefrom,
involving whole animals, including transgenic animals, and not covered by Sections III-D-1 or III-D-4-a, may be
conducted at the appropriate containment determined by the IBC.
III-D-4-c-1
Experiments involving the generation of transgenic rodents that require BSL1 containment are described under
Section III-E-3.
III-D-4-c-2
Purchase or transfer of transgenic rodents is exempt from the “NIH Guidelines” under Section III-F.
III-D-5
Experiments involving whole plants. Experiments to genetically engineer plants by methods using recombinant or
synthetic nucleic acid molecules, to use plants for other experimental purposes, to propagate such plants, or to use
plants together with microorganisms or insects containing recombinant or synthetic nucleic acid molecules , may
be conducted under the containment conditions described in the “NIH Guidelines”, Section III-D-5a through
Section III-D-5e.
III-D-6
Experiments involving more than 10 liters of culture. The appropriate containment will be decided by the IBC.
Where appropriate, Appendix K of the “NIH Guidelines” will be used to determine containment conditions.
III-D-7
Experiments Involving Influenza Viruses. Experiments with influenza viruses generated by recombinant methods
(e.g., generation by reverse genetics of chimeric viruses with reassorted segments, introduction of specific
mutations) shall be conducted at the biosafety level containment corresponding to the Risk Group of the virus that
was the source of the majority of segments in the recombinant virus (e.g., experiments with viruses containing a
majority of segments from a RG3 virus shall be conducted at BL3). Experiments with influenza viruses containing
genes or segments from 1918-1919 H1N1 (1918 H1N1), human H2N2 (1957-1968) and highly pathogenic avian
influenza H5N1 strains within the Goose/Guangdong/96-like H5 lineage (HPAI H5N1) shall be conducted at BL3
enhanced containment (see Appendix G-II-C-5, Biosafety Level 3 Enhanced for Research Involving Risk Group 3
Influenza Viruses) unless further noted in the Guidelines.
Section III-C Experiments that require IBC and Institutional Review Board (IRB) approvals, and RAC review before research
participant enrollment.
III-C-1
Experiments involving the deliberate transfer of (1) recombinant or synthetic nucleic acid molecules, or (2) DNA or
RNA derived from recombinant or synthetic nucleic acid molecules, into one or more human Research participants
Section III-B Experiments that require NIH/OBA and IBC approval before initiation.
13
III-B-1
Experiments involving the cloning of toxin molecules with LD50 of less than 100 nanograms per kilogram body
weight.
III-B-2
Experiments that have been Approved (under section III-A-1-a) as Major Actions under the NIH Guidelines (“me
too” experiments)
Section III-A Experiments that require Institutional Biosafety Committee (IBC) approval, Recombinant DNA Advisory Committee
(RAC) review, and NIH Director approval before initiation of experiments.
III-A-1-a
Deliberate transfer of a drug resistance trait to microorganisms that are not known to acquire the trait naturally, if
such acquisition could compromise the use of the drug to control disease agents in humans, veterinary medicine or
agriculture.
14