OREGON HEALTH & SCIENCE UNIVERSITY
Recombinant DNA (rDNA) Research Questionnaire (RDRQ)
All research involving rDNA or synthetic nucleic acid molecules must be reviewed by the IBC, regardless of funding.
For more information on these requirements, please see
NIH Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Molecules
For IBC Office Use: Registration # IBC-
Date:
(Please answer questions completely – Use additional space as necessary)
Principal Investigator (Last Name, First Name, Degree)
Telephone Number
Email Address
Mail Code
Laboratory Contact
Protocol Title
Are non-recombinant infectious agents or biologically-derived toxins also involved in this project?
If yes, you must also complete the Infectious Agent/Toxin Questionnaire.
Are select agents involved in this project?
See definition and FAQs at: http://www.ohsu.edu/xd/research/about/integrity/ibc/sa_faqs.cfm
Are agents, or genetic elements from agents on the Commerce Control List involved in this project?
Does research involved in this project require review by NIH-OBA?
See Fact Sheet on Research Requiring NIH OBA review (http://www.ohsu.edu/xd/about/services/integrity/policies/ibcpolicies.cfm#results)
Grant/Project Title(s) covered by this protocol (insert additional rows as needed)
Research personnel to work with
agent(s) on this project (including PI)*
Example: Gene X. Splicer
Role in project
-Production of adenoviral vectors and
performing injections
-Production of Lentiviral vectors
ePPQ#
Project
Sponsor
Yes
No
Yes
No
Yes
No
Yes
No
Check if
Funded
Check if
Pending
Description of training/experience
(relevant to role)
10 years
None to date
*If any personnel from a different lab are involved in any of the projects covered by the protocol, please clarify that lab’s
involvement in the project here and indicate at the end of this document whether this will be a Co-PI or single PI project:
For questions, contact the OHSU IBC analyst at (503) 494-7887 (option 1), the OHSU Central Campus Biosafety Officer, (503)
494-0655, or the OHSU West Campus Biosafety Officer, (503) 690-5312.
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Please answer all questions on this form. You may indicate “N/A” if a particular question is not applicable.
Please return signed original to Mail Code: L106-RI or email to: ibc@ohsu.edu
Summary of proposed work. Please use language appropriate for a scientific academician working in an unrelated
field.
A. Please provide a brief overview of the aims of the research protocol. [e.g. The lab has identified a
novel gene product that may be involved in neural plasticity and the functionality will be tested.]
B. How is recombinant DNA being used to address these aims? [e.g. lentiviral vectors are injected into
mouse brain for expression of gene of interest in neurons]
Please describe all non-exempt recombinant or synthetic DNA molecules to be used in this protocol. Please see
the FAQs on the IBC website for information on exempt and non-exempt nucleic acid molecules.
1. Please provide overview of rDNA molecules to be used, and whether in vivo uses are proposed
(use additional rows as necessary)
Vector type/Agent
Insert(s) or deletions
Host (cell type and/or species)
Example: lentivirus
Example: L. pneumophila
GFP
LP01,  WXYZ
Mice (in vivo)
human fibroblasts (in vitro)
Vector/Agent Detail
Please provide commercial product literature, a web link to specific information, a vector map, or a copy of any
journal articles describing construction of vector. Also state whether you will be producing the vectors, or
obtaining them commercially and/or from an OHSU Core facility:
2. A. Vectors to be used (plasmids, cosmids, phages, viruses)--specify type and strain, and give
description.
B. If plasmids are used in the packaging system for the viral vector, they must also be listed and
described.
C. If a packaging cell line is used for the viral vector, it must also be listed.
3. A. For viral vectors, describe how the vector differs from the original virus (in terms of
pathogenicity and genome size): [e.g., E1 & E3 genes deleted from adenoviral vector]
B. For recombinant infectious agents, describe how it differs from the wild type infectious agent
(in terms of pathogenicity, virulence, or immunogenicity): [e.g., attenuated Listeria monocytogenes]
4. List all drug resistance genes that are NOT for the purpose of plasmid propagation in E.coli or for
plasmid transfection in in vitro cell culture. Could any of these drug resistance genes compromise
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the use of the drug to control disease caused by this agent in humans, animals, or agriculture?
5. Will infectious virus particles or other infectious agents, either replication-deficient or replication
competent, be rescued, propagated or purified in your laboratory?
6. What is the host range of the viral vector or recombinant infectious agent?
7. Will there be surveillance for production of wild type or replication competent infectious agents?
Discuss.
DNA Insert(s) and Expression
NOTE: Additional review by NIH-OBA is required if: 1) DNA from smallpox (variola major or minor) or whitepox
is used; or 2) DNA encodes a gene for the biosynthesis of a toxin lethal for vertebrates with an LD50 of less
than 100 g/kg body weight. Additional information can be found on the Fact Sheets on Research requiring
NIH-OBA review and on Biological toxins on the IBC Policies and Forms webpage.
8. State DNA Source (species, tissue/cell, or microbiological agents).
9. Genetic elements contained in the inserted DNA sequences:
10. What gene products will be expressed or knocked down (shRNA or gene deletion) and what is
their function? If the gene product being expressed is from a Commerce Control Agent please
provide citations describing the function of the gene product.
11. Please describe potential human, animal, or plant health hazards of the gene product or its
transcribed or translated products (e.g., could it be oncogenic or toxic?). Please provide the
vertebrate LD50 for any genes encoding a biological toxin.
12. Would any of the gene inserts or deletions potentially increase the virulence of the recombinant
virus or recombinant pathogenic organism or allow the infectious agent to evade the host
immune response? If yes, discuss.
13. Will any gene be intentionally mutated? If yes, describe.
Host (cell type and/or species)
14. For in vitro experiments list host cells to be targeted (bacterial, eukaryotic, species).
15. Are viral sequences present in the host that could recombine with the vector and lead to
replication-competency for the recombinant construct, or has the host been previously infected
with another agent?
16. For in vivo experiments list host species and target organs or systems and describe the method of
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delivery, including the dose and the number of administrations. Provide IACUC number
associated with this project.
17. For whole animals, could there be an adverse physiological impact? Discuss.
18. For in vivo experiments, specifically discuss the potential for shedding of the agent from the
animal host.
19. For in vivo experiments, will fluids, tissues or cells be harvested from the animal? Briefly describe
harvested material and discuss if the material is potentially infectious or may contain replication
competent infectious material.
Biosafety
NOTE: Laboratories conducting research at biosafety level 2 or higher should have a lab specific biosafety manual.
Template biosafety manuals can be found on the OHSU IBC Policies and Forms webpage and may be useful
in addressing questions in the next section. If work is conducted at Biosafety Level 3 (BSL-3) the
laboratory specific manual must be submitted for IBC review and include standard operating procedures
outlining the laboratory specific protocols to be followed for work with BSL-3 agents. Contact the IBC if
guidance is needed.
20. Discuss the biohazard implications including potential exposure to staff and animal colonies.
Consider the consequences of an accidental exposure, e.g., spill, mucosal splash, inhalation, or
inoculation, which might occur during experimental handling. Describe protective
clothing/equipment that personnel are required to wear/use. Include other preventative
measures (e.g., vaccinations) if applicable.
21. Describe the containment facilities where these experiments (both in vitro and in vivo) will take
place. Please include all of the following: location, manufacturer, type, and certification date of
biosafety cabinets (tissue culture hoods) where these agents will be used.
22. Describe equipment to be used (e.g., flow cytometer, centrifuge, microscope), or transportation
to other locations that may require additional biosafety precautions.
23. Describe how materials containing rDNA or infectious agents (viral or other gene expression
vectors, transfected cell lines, infected tissues, etc.) will be disposed/discarded.
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Assurance
I will abide by the NIH Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Molecules the
CDC Guidelines for Biosafety in Microbiological and Biomedical Laboratories, and OHSU policies and procedures
for research involving rDNA molecules (http://www.ohsu.edu/xd/about/services/integrity/policies/ibcpolicies.cfm#results).
I will maintain a current record of any transfer of recombinant DNA or synthetic nucleic acid molecules or vectors
or host strains containing recombinant DNA or synthetic nucleic acid molecules, or infectious agents between
investigators at this or other institutions. I will follow IATA and CITES requirements and will ensure any laboratory
personnel have received the required training, when applicable, for shipment of biological materials (see
http://ozone.ohsu.edu/ehrs/mh/pages/bio/infsub.shtml). Additionally, I agree to comply with all U.S. export
control laws and OHSU policies and procedures when shipping materials internationally.
I agree that as Principal Investigator or Co-Principal Investigator it is my responsibility to make certain that prior
to engaging in research involving known or potential pathogens, all laboratory and support personnel are
properly trained in the practices and techniques required to ensure safety, and to supervise the safety
performance of those involved ensuring that the required safety practices and techniques are employed.
I agree to send a Project Modification Form to the Institutional Biosafety Committee if changes are made to the
recombinant DNA or synthetic nucleic acid molecule experiments described in this questionnaire.
PI/Head of lab:
Co-PI (if applicable):
The PI has overall responsibility for the project, including proper training and safety
supervision, and assuring that the IBC registration is up to date. Additional PI
responsibilities are posted on the IBC website.
Include a Co-PI when a project involves multiple lab heads contributing significantly to
the project and each share an equal responsibility in project oversight. If a Co-PI is
indicated, the responsibilities as indicated above apply to both individuals. The PI to
serve as primary contact should be indicated in the PI/Head of lab space.
Principal Investigator/Head of lab
Signature
Date
Co-Principal Investigator (if applicable)
Signature
Date
Co-Principal Investigator (if applicable)
Signature
Date
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EXPERIMENTS COVERED BY THE NIH GUIDELINES.
The Institutional Biosafety Committee may not authorize initiation of experiments which are not explicitly covered by the NIH Guidelines until NIH (with the
advice of the RAC when required) establishes the containment requirement (Section IV-B-2-b-(8)).
Section III-A. Experiments that Require Institutional Biosafety Committee Approval, RAC Review, and NIH Director Approval Before Initiation (See
Section IV-C-1-b-(1), Major Actions). Experiments considered as Major Actions under the NIH Guidelines cannot be initiated without submission of
relevant information on the proposed experiment to the Office of Biotechnology Activities, National Institutes of Health, the publication of the proposal in
the Federal Register for 15 days of comment, review by RAC, and specific approval by NIH. The containment conditions or stipulation requirements for
such experiments will be recommended by RAC and set by NIH at the time of approval. Such experiments require Institutional Biosafety Committee
approval before initiation. Specific experiments already approved are included in Appendix D, Major Actions Taken under the NIH Guidelines.
 Section III-A-1-a: The 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, will be reviewed by RAC. Consideration
should be given as to whether the drug resistance trait to be used in the experiment would render that microorganism resistant to the primary drug
available to and/or indicated for certain populations, for example children or pregnant women. At the request of an Institutional Biosafety Committee,
NIH/OBA will make a determination regarding whether a specific experiment involving the deliberate transfer of a drug resistance trait falls under
Section III-A-1-a. An IBC may also consult with NIH/OBA regarding experiments that do not meet the requirements of Section III-A-1-a but nonetheless
raise important public health issues. NIH/OBA will consult, as needed, with one or more experts, which may include the RAC.
Section III-B. Experiments that Require NIH/OBA and Institutional Biosafety Committee Approval Before Initiation
 Section III-B-1: Experiments Involving the Cloning of Toxin Molecules with LD50 of Less than 100 Nanograms per Kilogram Body Weight. Deliberate
formation of recombinant or synthetic nucleic acid molecules containing genes for the biosynthesis of toxin molecules lethal for vertebrates at an LD50
of less than 100 nanograms per kilogram body weight (e.g., microbial toxins such as the botulinum toxins, tetanus toxin, diphtheria toxin, and Shigella
dysenteriae neurotoxin). The containment for such experiments will be determined by NIH-OBA in consultation with Ad Hoc experts.
 Note: Additional information on regulated toxin experiments is found in Appendix F. Containment Conditions for genes coding the biosynthesis of
molecules toxic for vertebrates
 Section III-B-2. Experiments that have been Approved (Under Section III-1-a) as Major Actions under NIH Guidelines Upon receipt and review of an
application from the investigator, NIH/OBA may determine that a proposed experiment is equivalent to an experiment that has previously been
approved by the NIH Director as a Major Action, including experiments approved prior to implementation of these changes. An experiment will only be
considered equivalent if, as determined by NIH/OBA, there are no substantive differences and pertinent information has not emerged since submission
of the initial III-A-1-a experiment that would change the biosafety and public health considerations for the proposed experiments. If such a
determination is made by NIH/OBA, these experiments will not require review and approval under Section III-A.
Section III-C: Experiments Involving the Deliberate Transfer of Recombinant DNA, or DNA or RNA Derived from Recombinant DNA, into One or More
Human Research Participants Please refer to the Human Gene Transfer section of the IBC website for information on review requirements and applicable
forms.
Section III-D (experiments falling into any of the following categories below) The IBC reviews and approves all experiments in this category prior to their
initiation. Requests to decrease the level of containment specified for experiments in this section are Minor Actions under Section IV-C-1-b-(2)-(c)
requiring review by NIH-OBA.
 Section 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)
 Section III-D-1-a. Experiments involving the introduction of recombinant or synthetic nucleic acid molecules into Risk Group (RG) 2 agents will usually be
conducted at Biosafety Level (BL) 2 containment. Experiments with such agents will usually be conducted with whole animals at BL2 or BL2-N (Animals)
containment.
 Section III-D-1-b. Experiments involving the introduction of recombinant or synthetic nucleic acid molecules into RG 3 agents will usually be conducted
at BL3 containment. Experiments with such agents will usually be conducted with whole animals at BL3 or BL3-N containment.
 Section III-D-1-c. Experiments involving the introduction of recombinant or synthetic nucleic acid molecules into Risk Group 4 agents are not
permitted at OHSU.
 Section III-D-1-d. Experiments involving the introduction of recombinant or synthetic nucleic acid molecules into restricted agents are not permitted
at OHSU.
 Section 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
 Section 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
performed under BL2 containment. Experiments in which DNA from RG 4 agents is transferred into nonpathogenic prokaryotes or lower eukaryotes may
be performed under BL2 containment after demonstration that only a totally and irreversibly defective fraction of the agent's genome is present in a
given recombinant. In the absence of such a demonstration, BL4 containment shall be used. The Institutional Biosafety Committee may approve the
specific lowering of containment for particular experiments to BL1.
 Section III-D-2-b. Containment for experiments involving DNA from smallpox major, smallpox minor or whitepox transferred into nonpathogenic
prokaryotes or lower eukaryotes shall be determined by NIH/OBA.
 Section 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. Special care should be used in the evaluation of containment levels for experiments which are likely to either enhance the
pathogenicity (e.g., insertion of a host oncogene) or to extend the host range (e.g., introduction of novel control elements) of viral vectors under
conditions that permit a productive infection. In such cases, serious consideration should be given to increasing physical containment by at least one
level.
 Section III-D-3-a. Experiments involving the use of infectious or defective RG2 viruses (see Appendix B-II) in the presence of helper virus may be
conducted at BL2.
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 Section III-D-3-b. Experiments involving the use of infectious or defective RG 3 viruses (see Appendix B-III-D) in the presence of helper virus may be
conducted at BL3.
 Section III-D-3-c. Experiments involving the use of infectious or defective RG 4 viruses (see Appendix B-IV-D) in the presence of helper virus are not
permitted at OHSU.
 Section III-D-3-d. Experiments involving the use of infectious or defective restricted poxvirus (Section V-L) in the presence of helper virus are not
permitted at OHSU.
 Section III-D-3-e. Experiments involving the use of infectious or defective viruses in the presence of helper virus which are not covered in Section III-D-3a
through III-D-3-d may be conducted at BL1.
 Section III-D-4. Experiments Involving Whole Animals
 This section covers experiments involving whole animals 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 germ-line (transgenic animals) and experiments involving viable
recombinant or synthetic nucleic acid molecule-modified microorganisms tested on whole animals. For the latter, other than viruses which are only
vertically transmitted, the experiments may not be conducted at BL1-N containment. A minimum containment of BL2 or BL2-N is required.
 Section III-D-4-a. Recombinant or synthetic nucleic acid molecule, 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 BL1 or BL1-N 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 BL1 or BL1-N and appropriate to the organism under study.
 Section III-D-4-b. For 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 the appropriate containment shall be determined by the
Institutional Biosafety Committee.
 Section III-D-5. Experiments Involving Whole Plants. (Research not likely to be conducted at OHSU)
 Section III-D-6. Experiments Involving More than 10 Liters of Culture
 The appropriate containment will be decided by the Institutional Biosafety Committee. Where appropriate, Appendix K, Physical Containment for Large
Scale Uses of Organisms Containing Recombinant DNA Molecules, shall be used. Appendix K describes containment conditions Good Large Scale Practice
through BL3-Large Scale.
 Section III-D-7. Experiments Involving Influenza Viruses. Experiments with influenza viruses generated by recombinant or synthetic 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 or synthetic virus.
 Section III-D-7-a. Human H2N2 (1957-1968). Experiments with influenza viruses containing the H2 hemagglutinin (HA) segment shall be conducted at
BL3 enhanced. Experiments with the H2 HA gene in cold-adapted, live attenuated vaccine strains (e.g., A/Ann Arbor/6/60 H2N2) may be conducted at
BL2 containment provided segments with mutations conferring temperature sensitivity and attenuation are not altered in the recombinant or synthetic
virus. Experiments with RG 2 influenza viruses containing genes from human H2N2 other than the HA gene can be worked on at BL2.
 Section III-D-7-b. Highly Pathogenic Avian Influenza H5N1 strains within the Goose/Guangdong/96-like H5 lineage (HPAI H5N1). Experiments
involving influenza viruses containing a majority of genes and/or segments from a HPAI H5N1 influenza virus shall be conducted at BL3 enhanced
containment. Experiments involving influenza viruses containing a minority of genes and/or segments from a HPAI H5N1 influenza virus shall be
conducted at BL3 enhanced unless a risk assessment performed by the IBC determines that they can be conducted safely at BL2 and strain has been
excluded as a Select Agent. If appropriate containment level is uncertain, the risk assessment process is outlined in Section III-D-7-b.
 Section III-D-7-c. 1918 H1N1. Experiments involving influenza viruses containing any gene or segment from 1918 H1N1 shall be conducted at BL3
enhanced containment.
 Section III-D-7-d. Antiviral Susceptibility and Containment. The availability of antiviral drugs as preventive and therapeutic measures is an important
safeguard for experiments with 1918 H1N1, HPAI H5N1, and human H2N2 (1957-1968). If an influenza virus containing genes from one of these viruses
is resistant to both classes of current antiviral agents, adamantanes and neuraminidase inhibitors, higher containment may be required based on the
risk assessment considering transmissibility to humans, virulence, pandemic potential, alternative antiviral agents if available, etc.

Note: Experiments with 1918 H1N1, human H2N2 (1957-1968) or HPAI H5N1 that are designed to create resistance to neuraminidase inhibitors or
other effective antiviral agents (including investigational antiviral agents being developed for influenza) would be subject to Section III-A-1 (Major
Actions) and require RAC review and NIH Director approval, or if the agent is a Select Agent, the NIH will defer to the appropriate Federal agency
(HHS or USDA Select Agent Divisions).

Note: Details of BL3 Enhanced for Research Involving RG 3 Influenza viruses are found in Appendix G-II-C-5.
Section III-E (experiments falling into any of the following categories below)
 Experiments not included in categories A-D or F, are considered Category E. For example, experiments in which all components are derived from nonpathogenic prokaryotes and non-pathogenic lower eukaryotes.
 Section III-E-1. Experiments Involving the Formation of Recombinant or Synthetic Nucleic Acid Molecules Containing No More than Two-Thirds of the
Genome of any Eukaryotic Virus Recombinant or synthetic nucleic acid molecules containing no more than two-thirds 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 BL1 containment.) For
such experiments, it must be demonstrated that the cells lack helper virus for the specific Families of defective viruses being used. The DNA may
contain fragments of the genome of viruses from more than one Family but each fragment shall be less than two-thirds of a genome.
 Section III-E-2. Experiments Involving Whole Plants. (Research not likely to be conducted at OHSU)
 Section III-E-3. Experiments Involving Transgenic Rodents This section covers 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).
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