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
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
phone: (352)-392-1591
fax: (352)-392-3647
Section 1 – Basic Information
PI Name: / Title:Department: / Address/Box:
Office Phone: / Lab Phone: / Email:
Project Title:
Project Location: All Building(s): / All Room(s):
Sponsor:
Section 2 – General Project Information
2.1 Recombinant /Synthetic nucleic acid use:
YES / NOa. 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?)
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:
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?
2.2 Do your experiments have any of the following dual-use research of concern issues:
YES / NOa. 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?
If you checked yes to any of the above, please explain each in more detail:
2.3 Proposed Biosafety / Biocontainment Level for the project (as a whole or by parts):
Section 3 - Human Use
3.1 Will human subjects and/or human clinical specimens be used in this research? Yes No
If yes, have you received IRB approval?
No Date of intended submission:
Yes IRB#:
Approval pending – date submitted to IRB:
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. A description of the product:
1. Describe the derivation of the delivery vector system including the source (e.g., viral, bacterial, or plasmid vector); and modifications (e.g., deletions to attenuate or self-inactivate, encapsulation in any synthetic complex, changes to tropisms, etc.). Please reference any previous clinical experience with this vector or similar vectors.
2. Describe the genetic content of the transgene or nucleic acid delivered including the species source of the sequence and whether any modifications have been made (e.g. mutations, deletions, and truncations). What are the regulatory elements contained in the construct?
3. Describe any other material to be used in preparation of the agent (vector and transgene) that will be administered to the human research subject (e.g., helper virus, packaging cell line, carrier particles).
4. Describe the methods for replication-competent virus testing, if applicable.
5. Describe the intended ex vivo or in vivo target cells and transduction efficiency.
6. Describe the gene transfer agent delivery method.
b. Investigator Study Brochure
c. Investigator Study Protocol
d. Informed Consent document
e. Status of IRB review (IRB approval is not needed prior to IBC review – after is OK)
f. Confirmation that the NIH protocol registration process is complete
g. If the human gene transfer project was initiated and reviewed by the NIH prior to April 27, 2016, please submit the RAC approval letter, answers to Appendix M and items 3.2 b-e.
Please note that if UF is serving as the initial clinical trial site, all documentation described in Appendix M-I-A must be submitted to the NIH OSP before the IBC can approve the project. Required documentation includes a letter from the IBC detailing their assessment of whether public Recombinant DNA Advisory Committee (RAC) review is warranted. The IBC cannot approve any protocols until the NIH registration process is complete so please plan accordingly. For additional information regarding the registration and review process for human gene transfer protocols initiated on or after April 27, 2016, please see the following: http://osp.od.nih.gov/sites/default/files/HGT_FAQ%20sheet_508%20complaint.pdf.
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? Yes No
If yes,
a. Where will you create your transgenic animals?
With the help of the UF Mouse Models Core
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: Room: Other:
4.8 Where do you plan to perform animal procedures? Building: Room: Other:
Section 5 – Plant Use
5.1 Will you isolate nucleic acids from plants? Yes No
If yes, which plants? (list)
5.2 Will you transfer nucleic acid into plants? Yes No
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 Location:
Greenhouse Location:
Field Trial Location:
5.6 Are the plants regulated by the state or federal government as a noxious weed, invasive,
or exotic plant? Yes No
Section 6 – Insect or Arthropod Use
6.1 Will you isolate nucleic acids from insects or arthropods? Yes No
If yes, which nucleic acids? (list)
From which insect/arthropod(s)?
6.2 Will you transfer nucleic acid into insects/arthropods? Yes No
If yes, which nucleic acids (list)?
Into which insect/arthropod(s)?
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 Location:
Growth chamber Location:
Insectary Location:
Field Trial 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? Yes No
If yes, where will you obtain these?
7.2 Will primary human/non-human primate cells (collected directly from the human/primate) be used? Yes No
If yes, where will you obtain these?
Are these oncogenic/tumorigenic? Yes No
7.3 Will human or non-human primate cell lines be used? Yes No
Which cell lines? Where obtained (source or manufacturer, list)?
Are these oncogenic/tumorigenic? Yes No
Section 8 – Pathogen Use
8.1 Will you transfer genes/ nucleic acid from a pathogen into another organism/host? Yes No
To register more than 3 pathogens, complete an additional section 8.1 and attach to this form
Question Pathogen 1 Pathogen 2 Pathogen 3
Name of pathogenWhat 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? Yes No
To register more than 3 genes, complete an additional section 8.2 and attach to this form
Question Nucleic acid/gene 1 Nucleic acid/gene 2 Nucleic acid/gene 3
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? Yes No
To register more than 3 genes, complete an additional section 8.3 and attach to this form
Question Nucleic acid/gene 1 Nucleic acid/gene 2 Nucleic acid/gene 3
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?
Section 9 – Details about the recombinant or synthetic nucleic acids:
Detail recombinant/synthetic nucleic acids used in this project below.
9.1 Viral vectors:
To register more than 4 constructs/vectors, complete an additional section 9.1 and attach to this form
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)
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: / < 1/2
½ - 2/3
> 2/3
Other: / < 1/2
½ - 2/3
> 2/3
Other: / < 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:
Discuss safety features of the viral vectors you will be using (e.g. gene deletions, expression of packaging genes on multiple plasmids, self-inactivating 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
c. Assembled in my lab from components made/obtained elsewhere Yes No
Obtained from:
d. Received packaged virus Yes No
Obtained from:
e. Received transduced cells
Yes No
Obtained from: / a. Bought a commercial kit Yes No
Which:
b. Made all components in my lab Yes No
c. Assembled in my lab from components made/obtained elsewhere Yes No
Obtained from:
d. Received packaged virus Yes No
Obtained from:
e. Received transduced cells
Yes No
Obtained from: / a. Bought a commercial kit Yes No
Which:
b. Made all components in my lab Yes No
c. Assembled in my lab from components made/obtained elsewhere Yes No
Obtained from:
d. Received packaged virus Yes No
Obtained from:
e. Received transduced cells
Yes No
Obtained from: / a. Bought a commercial kit Yes No
Which:
b. Made all components in my lab Yes No
c. Assembled in my lab from components made/obtained elsewhere Yes No
Obtained from:
d. Received packaged virus Yes No
Obtained from:
e. Received transduced cells
Yes No
Obtained from:
Attach a map of all viral constructs at the end of the document
9.2 Plasmid vectors (transferable genetic elements, capable of autonomous replication within a suitable host)