BIOLOGICAL MATERIALS REGISTRATION FORM
Tufts University & Tufts Medical Center
Institutional Biosafety Committee (IBC)
Tel: 617-636-4109, Fax: 617-636-8354
E-mail: ibc-office@tufts.edu
Website: http://www.tufts.edu/central/research/IBC/
FOR IBC OFFICE USE ONLY
REGISTRATION #
APPROVAL DATE
EXPIRATION DATE
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 (NIH Guidelines), 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.
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INSTRUCTIONS for COMPLETING THIS FORM
Please type responses within the space/box provided and mark the checkboxes when appropriate.
Double-click on the checkboxes to mark your selections.
Submit the completed draft electronically to the IBC Office e-mail (ibc-office@tufts.edu).
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. CONTACT INFORMATION
PRINCIPAL INVESTIGATOR
ACADEMIC POSITION/TITLE
DEPARTMENT/DIVISION
LAB ADDRESS
DIRECT PHONE #
E-MAIL
LABORATORY CONTACT
DIRECT PHONE #
E-MAIL
DEGREE(S)
EMERGENCY #
FAX
DEGREE(S)
EMERGENCY #
II. LARGE SCALE RESEARCH
Will this research utilize viable organisms containing recombinant or synthetic nucleic acid molecules in culture
volumes of 10 liters or greater?
NO
YES
*Research utilizing volumes greater than or equal to 10 liters is defined as “Large Scale” and requires special permission. If
the project involves large scale work, please contact your Biosafety Officer for additional information.
III. PROJECT PERSONNEL
All listed personnel must complete mandatory IBC training. See “IBC Policy on Mandatory Biosafety Training”
and IBC website for information.
NAME
E-MAIL
PHONE #
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
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IV. BIOLOGICAL MATERIAL(S)
Provide the name(s) of each agent/material to be used. For recombinant DNA (rDNA) and synthetic nucleic acids,
mark the relevant Section of the NIH Guidelines under which it is described. Detailed definitions of each can be
found in Appendix A of this form. Identify the appropriate Biosafety Level (BSL) required for each agent proposed
according to the NIH Guidelines at http://oba.od.nih.gov/rdna/nih_guidelines_oba.html or the CDC/NIH Biosafety
in Microbiological and Biomedical Laboratories 5th edition at
http://www.cdc.gov/biosafety/publications/bmbl5/index.htm
If you have questions regarding these categorizations, the Biosafety Officer will help you during the pre-review.
A. RECOMBINANT AND/OR SYNTHETIC NUCLEIC ACID MOLECULES – List viral vectors here, if applicable.
Recombinant and synthetic nucleic acid molecules are defined as:
i) molecules that a) are constructed by joining nucleic acid molecules and b) can replicate in a living cell (i.e.
recombinant nucleic acids);
ii) nucleic acid molecules that are chemically or by other means synthesized or amplified, including those that
are chemically or otherwise modified but can base pair with naturally occurring nucleic acid molecules (i.e.
synthetic nucleic acids); or
iii) molecules that result from the replication of those described in (i) or (ii) above.
1. Name(s), Relevant Biosafety Level(s), and NIH Guidelines Categorization(s)
Name
Animal Biosafety Level
Biosafety Level
BSL-1
BSL-1
BSL-1
BSL-2
BSL-2
BSL-2
(containment/housing)
BSL-3
BSL-3
BSL-3
ABSL-1
ABSL-1
ABSL-1
ABSL-2
ABSL-2
ABSL-2
ABSL-3
ABSL-3
ABSL-3
Check all that apply below:
Section III-F: Experiments that are exempt from NIH Guidelines, but covered by local regulations.
Section III-E: Experiments that require IBC notice simultaneous with initiation.
Section III-D: Experiments that require IBC approval PRIOR TO initiation of experiments.
Section III-C: Experiments that require IBC and Institutional Review Board (IRB) approvals and Recombinant DNA
Advisory Committee (RAC) review before research participant enrollment.
Section III-B: Experiments that require NIH/OBA and IBC Approval BEFORE initiation of experiments.
Section III-A: Experiments that require IBC Approval, Recombinant DNA Advisory Committee (RAC) review, and NIH
Director Approval PRIOR TO initiation of experiments.
2. Is the agent attenuated or replication deficient?
NO
YES*
3. *If yes, please describe in detail the testing that will be done to confirm attenuation or replication
incompetence. Include the location of records. Please refer to the “Policy on Retroviral Replication
Competency Testing.”
NOTE: Insertion of oncogenes also requires testing (please describe below).
4. Please confirm that results of testing will be available for review.
YES
NO
B. INFECTIOUS AGENT(S) – Human source materials (e.g. blood or cell lines) should be reserved for Part F.
1. Name(s), Relevant Biosafety Level(s), and Pathology Information
Name
Biosafety Level
Animal Biosafety Level
(containment/housing)
Pathology of Agent
Human pathogen, Animal
pathogen, Both, Neither or
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N/A
BSL-1
BSL-1
BSL-1
BSL-2
BSL-2
BSL-2
BSL-3
BSL-3
BSL-3
ABSL-1
ABSL-1
ABSL-1
ABSL-2
ABSL-2
ABSL-2
ABSL-3
ABSL-3
ABSL-3
2. If a human pathogen, what is the infectious dose for a healthy human adult?
3. Is the agent attenuated or replication deficient?
NO
YES
4. If yes, please describe in detail the testing that will be done to confirm attenuation or replication
incompetence. Include the location of records.
5. Please confirm that results of testing will be available for review.
YES
NO
C. BIOLOGICAL TOXIN(S)
1. Name(s), Relevant Biosafety Level(s), and Toxicity Information
Name
Animal Biosafety Level
Biosafety Level
BSL-1
BSL-1
BSL-1
BSL-2
BSL-2
BSL-2
(containment/housing)
BSL-3
BSL-3
BSL-3
ABSL-1
ABSL-1
ABSL-1
ABSL-2
ABSL-2
ABSL-2
Toxicity of Agent
To humans, To animals, To
both, Neither or N/A
ABSL-3
ABSL-3
ABSL-3
2. What is the maximum quantity of toxin that will be present?
3. What is the LD50 of the toxin in humans?
4. What is the LD50 of the toxin in animal species used in this experiment?
D. SELECT AGENT(S) OR TOXIN(S) - To determine if your study falls within the Select Agent Rule, refer to
http://www.selectagents.gov/Select%20Agents%20and%20Toxins.html
1. Name(s), Relevant Biosafety Level(s), and Toxicity or Pathology Information
Name
Animal Biosafety Level
Biosafety Level
BSL-1
BSL-1
BSL-1
BSL-2
BSL-2
BSL-2
(containment/housing)
BSL-3
BSL-3
BSL-3
ABSL-1
ABSL-1
ABSL-1
ABSL-2
ABSL-2
ABSL-2
Pathology or Toxicity
Humans, Animals, Both,
Neither or N/A
ABSL-3
ABSL-3
ABSL-3
2. If a human pathogen, what is the infectious dose for a healthy human adult?
3. If a toxin, what is the maximum quantity 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?
E. GENETICALLY ENGINEERED ANIMALS - See “Policy on Genetically Engineered Animals” for detailed
information about the IBC review of genetic mutants and which type of
review is required.
1. Species and Relevant Biosafety Level(s)
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Animal Biosafety Level
Species
(containment/housing)
ABSL-1
ABSL-1
ABSL-2
ABSL-2
ABSL-3
ABSL-3
F. HUMAN SOURCE MATERIAL(S)
1. Employees of TUFTS MEDICAL CENTER are NOT required to be covered by the IBC for use of human source
materials. If you are a TMC employee, provide the appropriate confirmation below.
I confirm that all research with human source materials is conducted in the Tufts Medical Center facilities
and that annual Bloodborne Pathogen Training as a Tufts Medical Center employee is obtained.
OR
This does not apply to this registration.
2. Employees of TUFTS UNIVERSITY MUST register human source materials with the IBC. Provide the
information below.
HUMAN/NON-HUMAN PRIMATE (NHP) BLOOD, BODY FLUIDS, TISSUE, ORGANS
Animal Biosafety Level
Name
Biosafety Level
(containment/housing)
BSL-1
BSL-1
BSL-1
BSL-2
BSL-2
BSL-2
BSL-3
BSL-3
BSL-3
ABSL-1
ABSL-1
ABSL-1
ABSL-2
ABSL-2
ABSL-2
ABSL-3
ABSL-3
ABSL-3
HUMAN/NON-HUMAN PRIMATE (NHP) CELL LINES
Name
Animal Biosafety Level
Biosafety Level
BSL-1
BSL-1
BSL-1
BSL-2
BSL-2
BSL-2
(containment/housing)
BSL-3
BSL-3
BSL-3
ABSL-1
ABSL-1
ABSL-1
ABSL-2
ABSL-2
ABSL-2
ABSL-3
ABSL-3
ABSL-3
V. DUAL USE RESEARCH OF CONCERN (DURC)
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.
Enhances the harmful consequences of the agent or toxin
Disrupts immunity of 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, or involves agents or toxins with significant
potential for mass casualties or devastating effects to the economy, critical infrastructure, or public
confidence as listed in the “US Government Policy for Oversight of DURC,” Section (III.2)
None of the above applies
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VI. NON-TECHNICAL SUMMARY OF PROPOSED RESEARCH
Provide a BRIEF description of the goal of the research and how the agents are to be used in the box below. Lay
terminology must be used in place of field-specific terms and phrases.
*The blue field will expand as text is entered.
VII. EXPERIMENTAL DETAILS
In the following space, please summarize the intent of your proposed project. Provide enough information so that
the techniques and purposes of the experiments with the biohazardous agent(s) are clear. Indicate culture
volumes, maximum concentrations, and other agent-specific information. Identify at what stage of the
experiment the agent is inactivated. Be as concise as possible, using reasonably non-technical terms.
*The blue field will expand as text is entered.
VIII. EXPERIMENTAL MANIPULATION
Will the experiment(s) result in the acquisition of new characteristics such as enhanced virulence, infectivity, drug
resistance, or change in host range?
NO
YES*
*If yes, please explain.
IX. LOCATION(S)
List ALL laboratories where research is to be conducted and the corresponding biosafety level. Include cold/warm
rooms, equipment rooms, and location(s) of biosafety cabinets (BSC). For locations in the centralized animal
facilities, please indicate “DLAM” (Department of Laboratory Animal Medicine), “LAMS” (Laboratory Animal
Medicine Services),” or “CBU” (Comparative Biology Unit).
Room Purpose
Room Number for Labs
(Main Lab, Storage, Tissue Culture,
Procedure, etc.)
Biosafety Level
BSL-1
BSL-1
BSL-1
BSL-1
BSL-2
BSL-2
BSL-2
BSL-2
BSL-3
BSL-3
BSL-3
BSL-3
X. RISK ASSESSMENT
A. Hazardous processes used with agent. Check all that apply.
Centrifuge
Sharps
Animal Model
Sonication
Pipetting
Tissue harvesting
Tissue homogenization
Cell sorting (flow cytometry or other method)
Other: specify
B. Exposure route of agents. Check all that apply.
Ingestion
Percutaneous
Mucous Membrane
Inhalation
Other: (specify)
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.
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Personal Protective Equipment
Gloves
Lab Coat
Disposable Lab Gown
Disposable Booties
Tyvek Suit
N-95 Respirator
Surgical Mask
PAPR (Powered Air Purifying Respirator)
Safety Glasses
Other: (specify)
Engineering Controls
Biosafety Cabinet (BSC)
Other: (specify)
Agent(s) and Location of Use
Date of Certification
XI. EXPOSURE RESPONSE PLAN(S)
A. All labs working with biological agents must have an agent and lab specific exposure control plan. Please list
the titles of ALL Exposure Response Plan(s) associated with this registration. Many plans are available for
download on the IBC website. If the one you need is not there, please work with your Biosafety Officer to
create one.
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?
XII. DECONTAMINATION
A. What chemical and/or thermal processes will be used for decontamination?
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. Animal bedding:
If handled by DLAM/LAMS/CBU, please check the box:
Per standard ABSL-appropriate DLAM/LAMS/CBU procedures
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5. Animal cages:
If handled by DLAM/LAMS/CBU, please check the box:
Per standard ABSL-appropriate DLAM/LAMS/CBU procedures
6. Animal carcasses:
If handled by DLAM/LAMS/CBU, please check the box:
Per standard ABSL-appropriate DLAM/LAMS/CBU procedures
XIII. USE OF RECOMBINANT OR SYNTHETIC NUCLEIC ACID MOLECULES
Please answer the following questions in the space provided. Please write “N/A” for all items that do not apply.
A. SOURCE OF GENE, GENE FAMILY, INSERT, OR CLONE
1. Name the specific DNA/RNA source or probe (human, species of animal, plant, etc.)
2. What is the percent of viral genome in the construct?
3. What is the percent of synthetic DNA in the construct?
4. What is being expressed? What effect will be seen in the cell, animal etc.?
5. Provide information on any sequences that code for toxins.
B. VECTORS AND HOST CELLS
1. Identify the cloning/expression/transfection vectors used.
2. Identify the recipient bacterial strains and recipient host cell lines (human, species of animal, plant, etc.).
Provide a restriction map of vector unless this is a commercially available vector. If commercially available,
please indicate vendor.
3. Describe the location and type of promoters and other control sequences.
4. What is the percent of viral genome in the construct?
5. If using viral vectors, indicate packaging cell lines and assay system used to measure helper virus titer or titer of
replication competent virus (background) generated. Include host range of packaged viral vector (If using
retroviral or lentiviral vectors, additional requirements may apply).
6. What is the percent of synthetic DNA in the construct?
C. USE OF RECOMBINANT OR SYNTHETIC DNA IN ANIMALS, PLANTS, OR INSECTS
For genetically engineered rodent breeding at ABSL-1, information should be provided in the IACUC form only. See
“IBC Policy on Genetically Engineered Mutants.”
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.
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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. Provide location information for insects or plants. Include building and room number.
XIV. PROCEDURES WITH LIVE ANIMALS
Complete this section ONLY if the biological material(s) listed on this registration will be administered to animals.
Please provide a response for all items. If a question does not apply, please write “N/A.”
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. Location(s) of administration (building and lab room # OR DLAM/LAMS/CBU):
6. Will administration(s) be done inside a biosafety cabinet?
Yes
No
7. Will the biological agent or hazardous metabolite be excreted by urine, feces or wound drainage? Provide
duration, if applicable.
B. HOUSING/HANDLING INSIDE AND OUTSIDE OF CENTRALIZED FACILITIES
1. Will animals be removed from the centralized facilities?
Yes
No
a. If yes, will animals be returned to DLAM/LAMS/CBU post administration of the agent(s)?
No
Yes
b. Please indicate the Animal Biosafety Level necessary in the centralized facilities for the animals upon return:
ABSL-1
ABSL-2
2. Will a DLAM/LAMS/CBU biohazard cage card be used to identify the agent used?
a. If no, will an alternative method be used to identify the biohazardous agent?
b. Describe alternate method, if applicable.
Yes
No
No
Yes
3. Will handling and disposal of bedding, cages, and animal carcasses be done by DLAM/LAMS/CBU staff in
accordance with standard ABSL-appropriate DLAM/LAMS/CBU procedures?
Yes
No
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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.
OTHER HELPFUL LINKS
Tufts University EHS Website
Pathogen Safety Data Sheets
Biosafety in Microbiological and
Biomedical Laboratories (BMBL)
NIH Guidelines for Recombinant and
Synthetic Nucleic Acid Molecules
Tufts OVPR IBC Website
http://publicsafety.tufts.edu/ehs
http://www.phac-aspc.gc.ca/lab-bio/res/psds-ftss/index-eng.php#menu
http://www.cdc.gov/biosafety/publications/bmbl5/index.htm
http://oba.od.nih.gov/rdna/nih_guidelines_oba.html
http://www.tufts.edu/central/research/IBC/index.htm
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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. Mark the box next to the category that you chose.
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
Thosethat 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
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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.
Check 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 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.
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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 nontransgenic 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 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, IIIC, 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
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therefrom, into the germ-line (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.
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.
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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 IIID-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 IIIF.
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.
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III-B-1
III-B-2
Experiments involving the cloning of toxin molecules with LD50 of less than 100 nanograms per
kilogram body weight.
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.
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