Recombinant or Synthetic Nucleic Acid Research Registration
1) All projects involving recombinant nucleic acids must be registered. Projects involving synthetic nucleic acids or organisms or cells that contain synthetic nucleic acids must be registered provided that the synthetic nucleic acid is either a) designed to integrate into DNA b) replication competent or able to replicate in a living cell or c) codes for a vertebrate toxin with an LD50 of <100 nanograms/kilogram.
2) Registration & oversight of these projects is in accordance with the NIH GUIDELINES FOR RESEARCH INVOLVING
RECOMBINANT OR SYNTHETIC NUCLEIC ACID MOLECULES . In the context of the Guidelines , these molecules are defined as either: (i) molecules that a) are constructed by joining nucleic acid molecules and b) that 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. The Guidelines classify these experiments based on their potential hazard(s) to research staff, environment, and/or public health.
3) Projects must be approved BEFORE any research is initiated.
Important sections of the NIH Guidelines include:
Section III - Experiments covered by the NIH Guidelines (page 15)
Section IV B-7- Responsibilities of the PI (page 29)
Appendix B - Classification of Human Etiologic Agents on the Basis of Hazard (page 39)
Appendix G - Physical Containment; Biosafety Levels 1 –4 (page 71)
Appendix P - Containment for Recombinant or Synthetic Nucleic Acid Research Involving Plants (page 113)
Appendix Q - Containment for Recombinant or Synthetic Nucleic Acid Research Involving Animals (page 124)
Project submissions are reviewed by the Biosafety Office; non-exempt projects are then forwarded to the full Institutional Biosafety
Committee (IBC) for review, comment, and approval. The IBC is composed of scientists that may not be experts in your particular field of research . Please tailor your responses on the registration form accordingly. We must obtain sufficient information to determine required containment level, facilities, procedures, practices, and expertise/training necessary for the safe conduct of the project, therefore please be thorough.
Insufficient information/incomplete forms will delay the approval process and the form will be returned to you for revision. Please utilize the fillable document and type the form; hand written forms are not accepted.
If you have any questions, please contact the Biosafety Office at 392-1591 or bso@ehs.ufl.edu
The Biosafety in Microbiological and Biomedical Laboratories (5th Edition) is a valuable resource for details on containment, risk assessment, agent summary statements, etc. and can be found at: http://www.cdc.gov/biosafety/publications/bmbl5/index.htm

University of Florida
Environmental Health & Safety
Biological Safety Office bso@ehs.ufl.edu
phone: (352)-392-1591 fax: (352)-392-3647
Section 1 – Basic Information
PI Name:
Department:
Recombinant or Synthetic Nucleic Acid Registration
#RD -
Title:
Address/Box:
Email:
Project Title:
Project Location: All Building(s): All Room(s):
Sponsor:
Section 2 – General Project Information
2.1 Recombinant /Synthetic nucleic acid use: a.
Will proteins or regulatory RNAs be expressed? b.
Is the source of the DNA/nucleic acids to be used associated with alterations of normal mammalian cell cycle or cell growth (i.e. a potentially oncogenic or tumorigenic gene?)
YES NO
If yes, explain: c.
What is the maximum total volume of cultured recombinant/synthetic material being used at any one time? d.
Will your experiments be conducted outside of containment (e.g. outside the facility, field trial)?
If yes, explain: e.
Will you use any nucleic acid that codes for a vertebrate toxin with an LD50 of <100 nanograms/kilogram ?
If yes, describe:
2.2 Do your experiments have any of the following dual-use research of concern issues: f.
Will you use any nucleic acid designed to integrate into DNA? g.
Will you use any nucleic acid that is replication competent or able to replicate in a living cell? a.
Transfer drug, antibiotic, vaccine, or chemical resistance genes? b.
Alter genes associated with an agent’s pathogenicity (ability to cause disease)? c.
Alter genes associated with an agent’s virulence (extent or severity of disease)? d.
Alter genes associated with an agent’s transmissibility? e.
Enable evasion of diagnostic/detection modalities? f.
Alter the host range or tropism of a pathogen? g.
Change an organism’s normal ability to survive or be disseminated? h.
Enhances the susceptibility of a host/host population to a pathogen or toxin? i.
Reconstitutes an eradicated/extinct pathogen?
YES NO
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Section 3 - Human Use
3.1 Will human subjects and/or human clinical specimens be used in this research?
If yes, have you received IRB approval?
No Date of intended submission:
Yes IRB#:
Approval pending – date submitted to IRB:
Yes No
3.2 Human gene transfer projects involving Recombinant DNA, or DNA or RNA Derived from Recombinant DNA, into One or
More Human Research Participants will also require the following be submitted with this registration: a.
RAC approval letter or relevant RAC correspondence. See http://oba.od.nih.gov/rdna/rdna_faq_list.html
for more info b.
NIH Guidelines Appendix M filled out, a.k.a. “points to consider” : page 99 http://oba.od.nih.gov/oba/rac/Guidelines/NIH_Guidelines.pdf
c.
Investigator Study Brochure d.
Investigator Study Protocol e.
Informed Consent document f.
Status of IRB review (IRB approval is not needed prior to IBC review – after is OK)
The IBC requests that all human gene therapy trials go through a UF IRB.
(The WIRB can be the IRB of record, but the UF IRB reviews/comments before sending to WIRB )
Section 4 - Animal Use
4.1 Will your experiments involve the use of/introducing nucleic acid into animals? Yes No
4.2 Please list the animals you will be using:
4.3 How will the nucleic acid be transferred into the animal (e.g. viral vector, plasmid injection, transduced cells)?
4.4
Route of administration? Check all that apply.
Intravenous Intraperitoneal Subcutaneous Intracerebroventricular Intramuscular
Intranasal Other:
4.5 Will you create transgenics/knockouts?
If yes, a. Where will you create your transgenic animals?
With the help of the UF Mouse Models Core
Yes No
Within your own laboratory b. What method will be used to create your transgenic animals?
Microinjection of gene construct into pronuclear fertilized oocytes
Insertion of gene construct into embryonic stem cells that are microinjected into oocytes
Vector-mediated transfer of construct to embryonic stem cells for microinjection into oocytes
Other:
4.6 Has this project received approval from the UF IACUC?
No Date of intended submission:
Yes IACUC #:
Approval pending – date submitted to IACUC:
4.7 Where do you plan to house your animals? Building:
4.8 Where do you plan to perform animal procedures? Building:
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Room:
Room:
Other:
Other:
Recombinant/Synthetic Nucleic Acid ver 09/13

Section 5 – Plant Use
5.1 Will you isolate nucleic acids from plants?
If yes, which plants? (list)
5.2 Will you transfer nucleic acid into plants?
5.3 What type of plant(s) host?
5.4 How will you introduce nucleic acid into the plant?
5.5 Where will genetically modified plants be kept?
Laboratory Location:
Growth chamber
Greenhouse
Location:
Location:
Yes
Yes
No
No
Field Trial Location:
5.6 Are the plants regulated by the state or federal government as a noxious weed, invasive, or exotic plant?
Section 6 – Insect or Arthropod Use
6.1 Will you isolate nucleic acids from insects or arthropods?
If yes, which nucleic acids? (list)
From which insect/arthropod(s)?
6.2 Will you transfer nucleic acid into insects/arthropods?
If yes, which nucleic acids (list)?
Into which insect/arthropod(s)?
Yes
Yes
Yes
No
No
No
6.3 Is this insect regulated as a plant pest, exotic, or invasive species by the state or federal government? Yes No
6.4 How will you introduce nucleic acid into the insect/arthropod?
6.5 Where will the insects/arthropods be kept?
Laboratory
Growth chamber
Location:
Location:
Insectary
Field Trial
Location:
Location:
Section 7 – Use of human or non-human primate tissue, fluids, or cell lines
7.1 Will you use human or non-human primate tissues or fluids on this project?
If yes, where will you obtain these?
Yes No
7.2 Will primary human/non-human primate cells (collected directly from the human/primate) be used?
If yes, where will you obtain these?
Are these oncogenic/tumorigenic?
Yes No
Are these oncogenic/tumorigenic? Yes No
7.3 Will human or non-human primate cell lines be used?
Which cell lines?
Yes
Where obtained (source or manufacturer, list)?
No
Yes No
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Section 8 – Pathogen Use
8.1 Will you transfer genes/ nucleic acid from a pathogen into another organism/host?
If yes:
Question Pathogen 1 Pathogen 2
Name of pathogen
What pathogen genes/nucleic acids?
Gene/nucleic acid function?
Transferred into which host(s)?
8.2 Will you add genes /nucleic acids to a pathogenic organism
If yes:
Question Nucleic acid/gene 1 Nucleic acid/gene 2
Gene name?
Gene/nucleic acid function?
Gene/nucleic acid is from what organism?
Transferred into which pathogen(s)?
Is host range, pathogenicity, virulence, or antibiotic resistance of pathogen changed?
If yes, describe?
8.3 Will you delete genes /nucleic acids from a pathogenic organism
If yes:
Question Nucleic acid/gene 1 Nucleic acid/gene 2
Gene name?
Gene/nucleic acid function?
Deleted from what pathogen(s)?
Is host range, pathogenicity, virulence, or antibiotic resistance of pathogen changed?
If yes, describe?
Yes No
Pathogen 3
Yes No
Nucleic acid/gene 3
Yes No
Nucleic acid/gene 3
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Section 9 – Details about the recombinant or synthetic nucleic acids:
Detail recombinant/synthetic nucleic acids used in this project below. To register more than 4 constructs/vectors per table, print and attach additional pages as needed.
9.1 Viral vectors:
QUESTION Vector 1 Vector 2 Vector 3 Vector 4
Type of virus that makes up the vector system?
Name of vector construct?
Source of nucleic acid to be inserted?
Name of inserted nucleic acid?
Function of inserted nucleic acid?
Discuss risks associated with this inserted nucleic acid if it were accidentally transferred to an unintended host
(e.g. is it oncogenic, immunogenic, toxic, immune suppressive, angiogenic, an allergen, etc)
Is insertional mutagenesis a concern?
Why or why not?
List any key expression control elements
(amphotropic envelope gene, human promoter, etc.)
List ALL host(s) the vector will be transferred into. Include genus/species where applicable (e.g. packaging cell line, cell lines, prokaryotes, plants, animals, insects, etc)
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How will it be transferred to each host (transfection, viral transduction, transformation, infection, particle bombardment, injection, etc)?
How much will be transferred to each host
(concentration/titer and volume)?
What percentage of the viral genome will be transferred to the host(s)?
< 1/2
½ - 2/3
> 2/3
Other:
Is the virus capable of integrating into the host genome?
Is the virus replication defective?
If yes, describe
If no, provide justification for the use of a replication competent virus
Will you use defective viral vector in the presence of helper virus?
If yes, specify which:
List all helper plasmids used to produce recombinant virus
Has the vector preparation been tested
(e.g. by commercial vendor, another PI) or will it be tested in your lab for the presence of replication competent virus (RCV)?
If yes, describe the method used for testing and attach data as a separate sheet if available:
< 1/2
½ - 2/3
> 2/3
Other:
< 1/2
½ - 2/3
> 2/3
Other:
< 1/2
½ - 2/3
> 2/3
Other:
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Discuss safety features of the viral vectors you will be using (e.g. gene deletions, expression of packaging genes on multiple plasmids, selfinactivating long terminal repeats, limited tissue tropism).
How did you obtain the viral vector? a.
Bought a commercial kit Yes No
Which: b.
Made all components in my lab
Yes No a.
Bought a commercial kit Yes No
Which: b.
Made all components in my lab
Yes No a.
Bought a commercial kit Yes No
Which: b.
Made all components in my lab
Yes No a.
Bought a commercial kit Yes No
Which: b.
Made all components in my lab
Yes No c.
Assembled in my lab from components c.
Assembled in my lab from components c.
Assembled in my lab from components c.
Assembled in my lab from components made/obtained elsewhere
Yes No
Obtained from: made/obtained elsewhere
Yes No
Obtained from: made/obtained elsewhere
Yes No
Obtained from: made/obtained elsewhere
Yes No
Obtained from: d.
Received packaged virus
Yes No d.
Received packaged virus
Yes No d.
Received packaged virus
Yes No d.
Received packaged virus
Yes No
Obtained from: e.
Received transduced
Obtained from: e.
Received transduced
Obtained from: e.
Received transduced
Obtained from: e.
Received transduced cells
Yes No
Obtained from: cells
Yes No
Obtained from: cells
Yes No
Obtained from: cells
Yes No
Obtained from:
Attach a map of all viral constructs at the end of the document
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9.2 Plasmid vectors (transferable genetic elements, capable of autonomous replication within a suitable host) :
QUESTION Vector 1 Vector 2 Vector 3 Vector 4
Name of plasmid construct?
Name of inserted nucleic acid?
Source of nucleic acid inserted?
Function of inserted nucleic acid?
Discuss risks associated with this inserted nucleic acid if it were accidentally transferred to an unintended host
(e.g. oncogenic, immunogenic, toxic, immune suppressive, angiogenic, an allergen, etc)
List any key expression control elements (human promoter, enhancer, etc.)
List ALL host(s) the plasmid will be transferred into-list genus/species where applicable (e.g. cell lines, prokaryotes , plants, animals, insects)
How will it be transferred to each host?
(e.g. transformation, conjugation, electroporation, injection, particle bombardment, etc)
How much will be transferred to each host
(concentration and volume)?
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How did you obtain the plasmid vector: a.
Bought a commercial kit Yes No
Which: b.
Made all components a.
Bought a commercial kit Yes
Which:
No b.
Made all components a.
Bought a commercial kit Yes
Which:
No b.
Made all components a.
Bought a commercial kit Yes
Which:
No b.
Made all components in my lab
Yes No c.
Assembled in my lab from components made/obtained elsewhere
Yes No in my lab
Yes No c.
Assembled in my lab from components made/obtained elsewhere
Yes No in my lab
Yes No c.
Assembled in my lab from components made/obtained elsewhere
Yes No in my lab
Yes No c.
Assembled in my lab from components made/obtained elsewhere
Yes No
Obtained from: d.
Received cells already containing
Obtained from: d.
Received cells already containing
Obtained from: d.
Received cells already containing
Obtained from: d.
Received cells already containing
Yes No
Obtained from:
Yes No
Obtained from:
Yes No
Obtained from:
Yes No
Obtained from:
Attach a map of all vector constructs at the end of the document
9.3 siRNA/shRNA/miRNA/dsRNA
QUESTION siRNA 1 siRNA
siRNA 2 siRNA 3 siRNA 4
Description
Used with a plasmid or viral vector?
Transferred into what
(cell or animal)?
Target gene?
Where will siRNA be obtained from?
Yes No
Expected length of time of gene suppression
Is this able to suppress a beneficial gene?
Why or why not?
Could this have any detrimental off-target effects?
Why or why not?
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Yes No Yes No Yes No
Recombinant/Synthetic Nucleic Acid ver 09/13

QUESTION shRNA
Description
shRNA 1
Used with a plasmid or viral vector?
Transferred into what
(cell or animal)?
Yes No
Target gene?
Where will shRNA be obtained?
Expected length of time of gene suppression?
Is this able to suppress a beneficial gene?
Why or why not?
Could this have any detrimental off-target effects?
Why or why not?
QUESTION miRNA
miRNA 1
Description
Used with what plasmid or viral vector?
Yes No
Transferred into what
(cell or animal)?
Target gene?
Where will miRNA be obtained?
Expected length of time of gene suppression
Is this able to suppress a beneficial gene?
Why or why not?
Could this have any detrimental off-target effects?
Why or why not?
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shRNA 2
Yes No
shRNA 3 shRNA 4
Yes No Yes No
miRNA 2
Yes No
miRNA 3 miRNA 4
Yes No Yes No
Recombinant/Synthetic Nucleic Acid ver 09/13

QUESTION dsRNA
Description
dsRNA 1
Used with what plasmid or viral vector?
Transferred into what
(cell or animal)?
Yes No
Target gene?
dsRNA 2 dsRNA 3 dsRNA 4
Yes No Yes No
Where will dsRNA be obtained?
Expected length of time of gene suppression
Is this able to suppress a beneficial gene?
Why or why not?
Could this have any detrimental off-target effects?
Why or why not?
9.4 Other recombinant/synthetic nucleic acids used not described above:
Please provide a narrative or detail.
Yes No
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Section 10 – Research Description
10.1 Briefly describe the proposed work in lay terms. Your narrative must include: a.
brief introduction, b.
specific goal(s) of your experiment(s), c.
experimental approaches to be used with a focus on WHAT, WHY, and HOW recombinant or synthetic nucleic acids will be used, d.
the endpoints to be measured
The Biosafety Office/IBC does not review the scientific merit of the work but rather evaluates projects to ensure that they incorporate steps to minimize potential biohazard exposures and that biohazardous materials are disposed of in an appropriate manner. Provide sufficient information to ensure that the hazards and potential risks are easily identified.
Avoid using excess technical jargon and do not simply attach grant proposals, abstracts, manuscripts, or IACUC/lab protocols as a substitute.
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Section 11 – Risk Assessment
11.1 Discuss any risks associated with accidental exposure to or accidental environmental release of the recombinant or synthetic material or genetically modified organisms:
11.2 What section of the NIH Guidelines do these experiments fall under and what biosafety level would be appropriate for conducting these experiments ? (see The NIH Guidelines or the EH&S Biosafety Office NIH Guidelines Summary for assistance)
11.3 Describe how the following will be implemented to protect personnel from accidental exposure and/or prevent accidental environmental release: a. work practices: b. personal protective equipment (PPE): c. safety or containment equipment:
11.4
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Section 12 - Decontamination and Disposal
12.1 Which types of biological waste will be generated in your lab? Check all that apply.
Sharps
Recombinant /synthetic nucleic acid
Human pathogens
Animal pathogens
Plant pathogens
Human/primate blood, blood products, tissues, cultures, cells, or OPIM
Animal carcasses/tissues
Human remains/tissues
Mixed biological/chemical waste
Mixed biological/radioactive waste
12.2 Do you have access to an autoclave? Yes No
Building/room:
Proper function & testing monitored by (name):
Test method: Test frequency:
12.3 Do you have a copy of the biological waste disposal guidelines posted in the lab ( see http://www.ehs.ufl.edu/Bio/biowaste.htm#Policy and http://webfiles.ehs.ufl.edu/biomed_waste_disposal_guide.pdf
)? Yes No
12.4 Have all personnel been trained regarding proper biological waste disposal? Yes No
12.5 How will work surfaces be decontaminated?
12.6 How will solid waste be decontaminated and disposed of?
12.7 How will liquid waste be decontaminated and disposed of?
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Section 13 - Occupational Health and Training Information
Name and Title
Human/Non-human primate samples used
Blood
Tissues
Primary cell cultures
OPIM:
None
Blood
Tissues
Primary cell cultures
OPIM:
None
Blood
Tissues
Primary cell cultures
OPIM:
None
Blood
Tissues
Primary cell cultures
OPIM:
None
Blood
Tissues
Primary cell cultures
OPIM:
None
Blood
Tissues
Primary cell cultures
OPIM:
None
Vaccinations/Tests received
Hepatitis B
Vaccinia
TB screening
Serum banking
Respirator fit testing
Other:
Hepatitis B
Vaccinia
TB screening
Serum Banking
Respirator fit testing
Other:
Hepatitis B
Vaccinia
TB screening
Serum Banking
Respirator fit testing
Other:
Hepatitis B
Vaccinia
TB screening
Serum Banking
Respirator fit testing
Other:
Hepatitis B
Vaccinia
TB screening
Serum Banking
Respirator fit testing
Other:
Hepatitis B
Vaccinia
TB screening
Serum Banking
Respirator fit testing
Other:
Training Courses
Taken
Years experience working at:
BSL-1 BSL-2 BSL-3
BBP
BMW
Shipping &
Transport
Biosafety
BBP
BMW
Shipping &
Transport
Biosafety
BBP
BMW
Shipping &
Transport
Biosafety
BBP
BMW
Shipping &
Transport
Biosafety
BBP
BMW
Shipping &
Transport
Biosafety
BBP
BMW
Shipping &
Transport
Biosafety
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Section 14 – Project Personnel and PI Assurance
The undersigned individual(s) will be involved in the experimentation described above. They are familiar with and agree to abide by the current NIH Guidelines . ALL PARTICIPANT SIGNATURES REQUIRED.
Name (type or print) Title UF ID Signature Date
I attest to the fact that these individuals are properly trained in the area of recombinant /synthetic nucleic acid experimentation.
Furthermore, I agree to comply with the NIH requirements pertaining to handling, shipment, transfer, and disposal of recombinant/synthetic nucleic acid materials.
I am familiar with and agree to abide by the provisions of the current NIH Guidelines and other specific NIH instructions pertaining to the proposed project.
I understand that I must have EH&S or IBC approval before beginning this work.
I understand that changes to the project described above must be reported to EH&S Biosafety in advance.
I understand that associated IACUC or IRB approvals may be held pending EH&S or IBC approval of this work.
The information above is accurate and complete to the best of my knowledge.
Principal Investigator Date
E-mail ( bso@ehs.ufl.edu
), fax (352-392-3647), or mail (PO Box 112190) the completed form to the Biosafety office. Please see the calendar for IBC meeting dates and submission deadlines. Projects received after the 5pm deadline will be reviewed the following month.
Please include any construct/vector/plasmid maps as attachments.
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