University of South Carolina
Recombinant DNA Application
IBC-rDNA protocol #:
Final Instructions
1) Submit the application electronically via email attachment to: ibc@mailbox.sc.edu
2) Submit the Investigator’s NIH-style biosketch via email attachment
3) Fax a Signed copy of ONLY the first page to EHS FAX: 803-777-5275
SECTION A. INVESTIGATOR INFORMATION: It is the Principal Investigator’s responsibility to ensure all
personnel involved in this study are appropriately trained, and are provided the equipment necessary to perform at the
designated containment level.
A. 1. Investigator Assurance for Research Projects involving Recombinant DNA Molecules
a. I agree to conduct this project in accordance with the
policies of the University of South Carolina
Institutional Biosafety Committee (IBC), including all
requisite training of students, staff and other
professionals participating in this project.
e. If funded by an extramural source, I assure that this
application accurately reflects all procedures involving
recombinant DNA as described in the grant proposal
to the funding agency.
f.
b. I have consulted Section IV-B-7 of the NIH
Guidelines describing the responsibilities of the
Principal Investigator and hereby agree to comply
fully with all provisions of the NIH Guidelines
c. I understand that I am responsible for assuring that my
project areas are in compliance with local, state and
federal environmental laws and regulations.
d. I understand that all changes in the research protocol
(including changes in the source of DNA, hostvector systems, dosage ranges, laboratory room
changes, etc.) or project participants must be
reported to the IBC and all other university
regulatory offices in connection with this protocol.
The information within this application is accurate to
the best of my knowledge.
g. I understand that all protocols must be resubmitted for
committee review after a term of three years.
h. By the submission and acceptance of this signed
document by the IBC, I am in agreement with the
statements a-h (above).
The IBC in conjunction with the EHS Office
reserve the right to conduct inspections of the
research
facilities
at
any
time.
Investigator’s name typed:
Investigator’s signature
February 12, 2016
Project Title(s)
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Information for Applicants
Registration Term: IBC-rDNA protocols will be valid for a period of three years, after which time a full renewal must
be submitted to the committee for review and approval of continuing projects.
Modifications: Modifications to approved protocols must be approved by the IBC prior to implementation.
Modifications that involve a change in the assigned biosafety level of the project or risk group designation (for example,
in addition to plasmid vectors, the modification will add adenoviral vectors, moving from BSL-1 and risk group 1 to BSL2, risk group 2) must be submitted as either a full renewal or a new application. The IBC reserves the right to request a
complete application for modifications that do not qualify as minor modifications.
Termination: Protocols that are not renewed expire the day following the expiration date and are automatically
terminated. A termination letter is distributed to the investigator and other compliance offices/divisions as appropriate.
Terminated protocols may not be “re-started”. A new application must be submitted.
Containment determination and issues to consider: In determining the appropriate containment for a project, it is
important to consider factors that may raise concerns regarding the materials or agent(s) used.
Factors to consider in determining the level of containment include: 1) virulence, 2) pathogenicity, 3) infectious dose, 4)
environmental stability/instability, 5) route of spread/infection, 6) communicability/pathogenicity, 7) safety
procedures/operations, 8) quantity of agent(s), 9) availability of vaccine or treatment and, 10) any gene product effects
such as: toxicity, physiological activity, or allergenicity.1
Careful consideration should be given to the type of manipulation planned for some higher risk group agents. Any strain
that is known to be more hazardous than the parent (wild-type) strain should be considered for handling at a higher
containment level.2
Investigators should additionally consider any potential for unintended adverse events and/or potential for misuse of the
research. Special consideration should be given to any experimental paradigms that: 3
 Would demonstrate how to render a vaccine ineffective and thus potentially jeopardize public health
 Would confer resistance to therapeutically useful antibiotics or antiviral agents
 Would enhance the virulence of a pathogen or render a non-pathogen virulent
 Would increase transmissibility of a pathogen
 Would alter the host range of a pathogen
 Would enable the evasion of diagnostic/detection modalities
 Would enable the weaponization of a biological agent or toxin
1,2
NIH Guideline for Research Involving Recombinant DNA Molecules (NIH Guidelines), II-A-3, 2002
National Research Council, Biotechnology Research In An Age of Terrorism, National Academies Press, 2004
3
Application instructions: Place all responses in the UN-shaded areas. Do not type in the gray-shaded sections. Do not
leave any blanks, unless instructed to do so. Incomplete applications will be returned.
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A. 2. Investigator information
Investigator name
Professional title (e.g. Associate Professor)
Department/Division
Office room & building
Office telephone
Office facsimile
Laboratory room & building
E-mail address
A. 3. Alternate contact information
Alternate contact name (e.g. research
coordinator, technician, collaborator, etc.)
Title
Location; room and building
Telephone
Facsimile
E-mail address
SECTION B.
PROJECT INFORMATION:
B.1. Project submission:
Mark the type of submission for this application in the un-shaded box↓
Complete ALL Sections of the application, provide NIH biosketch
New rDNA project
Required every 3 years; Complete ALL Sections of the application, provide
NIH biosketch. If there are changes in the BSL a full renewal is required.
Modification type 1 Only changes which are clerical in nature: Title changes, funding, provide NIH biosketch
Complete Sections: A1, A2, A3, B1, B2, and other Sections as appropriate
Modification type 2 New Vectors: no change in BSL, no change in risk groups, provide NIH biosketch
Complete Sections: A1, A2, A3, B1, B2, and other Sections as appropriate
Full renewal of existing protocol
B.2. Previously approved IBC-rDNA protocol
Provide the IBC-rDNA protocol reference number of the protocol being renewed, modified or updated:
EHS Office Staff Use Only
IBC-rDNA protocol number:
Receipt date:
EHS-F-012
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B.3. Project summary
Describe the experimental procedures involving recombinant DNA materials. Use non-technical terminology to
enable IBC community representatives to understand your project.
Include in this summary the following
information:
1) The nature and purpose of the research
2) The procedures and techniques to be used in the project
The summary should not be copied directly from a grant application unless it is written in lay language and should
be limited to one page. The IBC reserves the right to return any proposals that do not meet the above conditions
or require extensive clarification or corrections prior to committee review.
Begin project summary here:
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B.4. Determination of use
NIH Guidelines
Please indicate all that apply by marking the check boxes
↓
references
a.
Using rDNA molecules for detection purposes (e.g. probes)
III-F
b.
Creating or using c-DNA/genomic libraries
III-E, III-F
c.
Cloning and vector construction in bacteria and yeasts
III-E, III-F
d.
III-B-1
g.
Using or cloning of toxin molecule genes (e.g. deliberate formation)
Using or cloning of genes from, or into a risk group 2, 3, 4 or restricted agent
(e.g. adenovirus, herpesvirus, retrovirus)
Transfer of a drug resistance trait into a risk group 2, 3, 4 or restricted agent
(e.g. adenovirus, herpesvirus, retrovirus)
Expression of rDNA products in cultured cells
h.
Propagating culture volumes exceeding 10 liters at one time
III-D-6
i.
Using virus(es) or viral vector(s)
III-D-3
j.
Using human cells/cell lines or tissues (e.g. human blood, 293 cell lines, CSF)
II-A-3
k.
Administration of rDNA product into humans (e.g. gene transfer)
III-C-1
l.
III-D-4
m.
Administration of rDNA material into animals (e.g. transformed cells, vectors)
Using animal cells/cell lines or tissues (e.g. tissue culture research)
n.
Using genetically modified animals in research (e.g. transgenic, knockout)
III-D-4-c(i)
o.
Using plants in research
III-D-5
e.
f.
III-D-1
III-D-2
III-A-1-a
III-E, III-F
II-A-3
III-E-1
Appendix C
SECTION C. BIOSAFETY INFORMATION:
Reference the NIH Guidelines Sections: II-A-1, IV-B-7-c, Appendix B and Appendix G
Biosafety level determination
Note: more than one biosafety level may apply to your project
1. Indicate the biosafety level(s) at which work is performed for this project by marking the un-shaded box(es)
 Low risk agents (generally risk group 1), special containment equipment not required
 Work is done on open bench tops
BSL-1
 Standard microbiological practices are observed
 Biohazard signs should be posted
BSL-2





Moderate risk agents (generally risk group 2), biosafety cabinets, restrictions to research areas
All BSL-1 containment and practices plus the following:
Laboratory access is restricted when experimental work is in progress
Personnel have specific training in handling of agents
Biological safety cabinets (BSC) or other physical containment devices are used for potential aerosol
generation procedures
 Biohazard signs must be posted
 Specific PPE (personnel protective equipment) and entrance requirements
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BSL-2+
BSL-3







Moderate-High risk agents (generally risk groups 2 or 3), BSL-2 containment with BSL-3 practices
All BSL-2 containment and practices plus the following:
Laboratory access is restricted
Personnel have specific training in handling of agents
All procedures are performed in biological safety cabinets (BSC)
Biohazard signs must be posted
Written safety policies provided by the investigator defining laboratory procedures, waste disposal,
disinfection and medical surveillance
 Centrifuge safety cups must be used
 NO current facilities exist at the University of South Carolina
 NO current facilities exist at the University of South Carolina
BSL-4
2. Indicate the risk groups (or class) of all material(s) used in this project by marking the un-shaded box
Risk Group 1 Agents are Not associated with disease in healthy adult humans.
Risk Group 2 Agents are associated with human disease that is rarely serious.
There are often preventive or therapeutic interventions available.
Risk Group 3 Agents are associated with serious or lethal human disease for which preventive or therapeutic
interventions MAY be available.
Risk Group 4 Agents are likely to cause serious or lethal human disease for which preventive or therapeutic
interventions are NOT USUALLY available.
Biosafety risks and training:
3. Is the highest proposed BSL lower than the risk group classification
for any of the described agents? (e.g. Use of single proteins from a risk
group 3 organism may qualify for work at BSL-1 or BSL-2)
(Mark “X” for Yes/No in un-shaded box)
If “Yes”, explain the rationale for the use of the lower BSL
4. Describe the potential biosafety risks of this research proposal.
Address the following:
a) whether agent(s) may be infectious to humans
b) the risks of accidental exposure to personnel
c) whether there is potential for airborne transmission of
agent(s)
d) precautions to be taken by personnel including any personal
protective equipment used and/or routine monitoring (e.g.
Tb testing)
e) disposal of agent(s); describe the specific methods of disposal
or inactivation of the agent or contaminated/infectious
material(s)
6. Describe the specific training and experience of the investigator.
Provide the years of experience with the agents in the application
and University of South Carolina training
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No
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Vectors, hosts, and recombinant DNA agents
 If desired, insert vector map(s) at the END of Section F
7. List all plasmid backbones used (e.g. pUC19, pGEM).
If none are used, state “None”
8. List any oligonucleotides used to manipulate gene function (e.g.
siRNA) or as adjuvants (e.g. CpG-containing DNA) either in cell
culture or in vivo.
If none are used, state “None”
9. List inserted DNA used; include the species from which the
insert is derived and what gene product is expressed.
If no inserts are used, state “None”
10. List known oncogenes, or inserts from above that have
oncogenic properties.
If none are used, state “None”
11. Is there any potential for increased virulence with insertion of DNA
into the vector or organism?
(Mark “X” for Yes/No in un-shaded box)
 If “Yes”, explain in detail
12. Are toxins to be expressed and released as part of this research?
(Mark “X” for Yes/No in un-shaded box)
 If “Yes”, describe the toxic product(s) (LD50 of <100 g/kg) that
could be produced/released and the containment precautions to
be used
13. List all bacterial and/or fungal agents used.
 If none are used, state “None”
14. List all eukaryotic cells, organisms, cell lines (including
human cell lines) or others.
 If none are used, state “None”
15. List other organisms (e.g. amoebas, nematodes).
 If none are used, state “None”
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Yes
No
Yes
No
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Viruses/viral vectors
16. Does this project involve the use of viruses or viral vectors?
(Mark “X” for Yes/No in un-shaded box)
Yes
No
 If “No”, skip to Section D
 If the virus or viral vector is a lentivirus (e.g. FIV, HIV, SIV, etc) complete questions 22-26
 For all other viruses or viral vectors, complete questions 17-20
 If desired, insert vector map(s) at the END of Section G
17. List viruses and/or viral vectors used:
 Specify the virus family and/or subfamily (e.g. herpesvirus,
oncogenic retrovirus, adenovirus, adeno-associated virus, etc.)
 State the species of origin for each virus or vector used
18. Is the virus/viral vector able to enter or infect human cells?
Yes
No
(Mark “X” for Yes/No in un-shaded box)
 If “Yes”, indicate whether it is a productive or limited infection,
and state whether infection can cause disease
19. Is a helper virus used in this project?
Yes
No
(Mark “X” for Yes/No in un-shaded box)
 If “Yes”, describe the helper virus used
20. Is the virus/viral vector replication-defective?
Yes
No
(Mark “X” for Yes/No in un-shaded box)
 If “No”, skip to Section D (unless also using Lentivirus/Lentiviral
vectors)
 If “Yes”, describe the deletions rendering it defective and complete
question 21
21. Has the replication-defective vector been tested for replication
Yes
No
competent virus?
(Mark “X” for Yes/No in un-shaded box)
 If “Yes”, provide details of the assay used
 If “No”, what is the likelihood of conversion to replicationcompetent virus?
Lentivirus/Lentiviral Vectors
22. List the specific virus or strain and species of origin.
(e.g. HIV, human; FIV, feline)
23. Is the lentivirus/lentiviral vector obtained from a commercial source?
(Mark “X” for Yes/No in un-shaded box)
 If “Yes”, provide the name of the commercial source (e.g. company
name)
 If “No”, provide the source of the lentivirus/lentiviral vector (e.g.
the name of the institution or individual supplying the material)
24. Is the lentivirus/lentiviral vector generated from a multi-component
system? (e.g. separate plasmids for packaging, envelope and gene transfer)
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(Mark “X” for Yes/No in un-shaded box)
 If “Yes”, describe the system used
25. Is the lentivirus/lentiviral vector pseudotyped (e.g. expressing a
different envelope gene)?
(Mark “X” for Yes/No in un-shaded box)
If “Yes”, provide whether the pseudotyping alters the host and cell
tropism
26. Is the lentivirus/lentiviral vector replication-defective?
(Mark “X” for Yes/No in un-shaded box)
 If “No”, skip to Section D
 If “Yes”, describe the deletions rendering it defective and complete
question 27
27. Has the replication-defective vector been tested for replicationcompetent virus?
(Mark “X” for Yes/No in un-shaded box)
 If “Yes”, provide details of the assay used
 If “No”, what is the likelihood of conversion to replicationcompetent virus?
SECTION D.
ANIMAL USE INFORMATION:
Yes
No
Yes
No
Yes
No
Reference NIH Guidelines II-A-1, Appendix B, III-E-3
Please visit the IACUC website for additional information
28. Does the work involve animal use?
(Mark “X” for Yes/No in un-shaded box)
 If “No” skip to Section E
29. Has an Institutional Animal Care and Use Committee (IACUC) application
been submitted?
(Mark “X” for Yes/No in un-shaded box)
 If “Yes” provide the IACUC protocol number to be linked to this rDNA
project (provide the temporary number or the date of submission, if the
IACUC number is not yet known)
Yes
30. Will animal tissues or cells be used in vitro?
(Mark “X” for Yes/No in un-shaded box)
 If “Yes”, explain
31. Will transgenic or gene targeted animals be used?
Yes
(Mark “X” for Yes/No in un-shaded box)
 If “Yes”, explain
32. What are the target cells/tissues/organs for the
recombinant materials?
33. Will recombinant agents be administered to live or intact animals? (viral
vectors, transfected cells, plasmids)
(Mark “X” for Yes/No in un-shaded box)
 If “Yes”, answer questions 34-38
 If “No”, skip to Section E
34. Do you anticipate that work with animal subjects will be conducted at a
different BSL than the in vitro portions of the study?
(Mark “X” for Yes/No in un-shaded box)
 If “Yes”, provide the BSL for work with or housing of animal subjects, and
explain the rationale or justification for the proposed BSL
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Yes
No
Yes
No
No
No
Yes
No
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No
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35. List each recombinant agent to be administered to animals
36. Provide the animal species (and strain if applicable) receiving the
recombinant DNA material; list each species to be used
37. Describe the route of administration for each recombinant agent
used
38. Provide the concentration and volume for each recombinant
agent to be administered
SECTION E.
HUMAN USE INFORMATION:
Reference NIH Guidelines III-B, III-C
39. Does work involve human cell lines (including cell lines such as 293T, HeLa),
human subjects, or unfixed human tissues or blood?
(Mark “X” for Yes/No in un-shaded box)
 If “No” skip to Section F
 If “Yes”, the cell lines used in this project must be listed under question 14
40. Has an Institutional Review Board (IRB) application been submitted?
(Mark “X” for Yes/No in un-shaded box)
 If “Yes” Provide the IRB protocol (preferred) or the IRB submission date
41. Will human tissues or primary cells be used in vitro?
(Mark “X” for Yes/No in un-shaded box)
 If “Yes”, describe the use of the tissues or cells
42. Is this a gene transfer proposal (recombinant materials administered to human
subjects)?
(Mark “X” for Yes/No in un-shaded box)
SECTION F.
Yes
No
Yes
No
Yes
No
Yes
No
FACILITIES INFORMATION:
Facilities: Locations where this project will be conducted
43. Provide the facilities information for ALL locations of this project, including the location of work involving
animals and human subjects, as applicable to your project.
 Describe the procedures performed in each location, for example, cell transfections, propagation of
plasmids, administration of viral vector into animals, animal housing, etc
 Provide the EHS approved biosafety level of the location. If a specific site is not currently EHS
approved, please state and explain
Room number and building
Describe procedures for this location
Location #1
Provide approved biosafety level
Location #2
Room number and building
Describe procedures for this location
Provide approved biosafety level
Location #3
Room number and building
Describe all procedures
Provide approved biosafety level
Location #4
Room number and building
Describe procedures for this location
Provide approved biosafety level
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Additional Materials
Insert additional materials in the box below if desired. For example, restriction map(s), host-vector diagrams, etc.
Note that the NIH Biosketch should be provided as a separate document.
Insert map(s)/diagrams here:
H EHS-F-012
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FOR IBC-rDNA USE ONLY
rDNA Protocol #
Instructions to Primary Reviewers:
When preparing comments/concerns for the investigator to address, reference the question number from the application.
Word comments carefully; the EHS Office staff will send comments verbatim to the investigator for response.
IBC-rDNA Committee Member Name:
2/12/16
IBC-rDNA protocol number:
Reviewer evaluation, summary or concerns:
Reviewer recommendation: (Mark an “X” in the blue-shaded box)
Approval; no revisions required. Current application is approvable
Approval pending; minor revisions required A revised application is required for approval
Additional information required; substantial revisions are needed or important information is missing from the
application
Defer to convened meeting; biosafety issues are identified which required full committee discussion
This form has been adapted from a University of Pittsburgh rDNA form.
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