HOW CAN WE FACILITATE UNDERSTANDING OF THE r-dna GUIDELINES TO THE SCIENTIFIC COMMUNITY? Esmeralda Meyer Kalpana Rengarajan Patty Olinger

Transcription

1 HOW CAN WE FACILITATE UNDERSTANDING OF THE r-dna GUIDELINES TO THE SCIENTIFIC COMMUNITY? Esmeralda Meyer Kalpana Rengarajan Patty Olinger October 13, 2015

2 Topics 1. Objective of the NIH Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Research (The NIH Guidelines) 2. Key Players involved in the application of the NIH Guidelines 3. Emory EHSO experience with reference documents 4. Why do we need a reference sheet for r-dna Classification? 5. Who will benefit? 6. What are the components of the document? 7. Plans for roll-out 8. Ongoing process

3 1. The NIH Guidelines Provides recommendations on how to conduct research with r-dna safely Revision in 2013 Section I: Oversight Section II: Risk Groups Section III: Classification of Experiments Section IV: Roles and responsibilities

4 2. Key Players involved in the application of the NIH Guidelines NIH Funded Institution Investigator Office of Biotechnology Activities Research Administration Biosafety Office & Institutional Biosafety Committee All Research Community from an institution which receives funding from NIH - Provides Guidance r-dna Guidelines - Oversight (Reporting agency) - Provides guidance -Reviews research -Ensures compliance -Informs the type of research to be conducted

5 What is r-dna? In the context of the NIH Guidelines, recombinant and synthetic nucleic acids are defined as: (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; or (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.

6 What type of review is required? In the context of the NIH Guidelines: (i) Section III-A: Institutional Biosafety Committee (IBC) approval + RAC + NIH Director approval before initiation (ii) Section III-B: NIH/OBA + IBC approval before initiation (iii) Section III-C: IBC + IRB + RAC review before research participant enrollment (iv) Section III-D: IBC approval before initiation (v) Section III-E: IBC notification simultaneous with initiation (vi) Section III-F: exempt from the NIH Guidelines

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9 4. Why do we need a reference sheet for r-dna? NIH Guidelines are comprehensive and not always easy to understand. Investigators must disclose not only the agents but details about the procedures. Providing examples for the guidelines facilitates applicability

10 5. Who will benefit from the use of a reference document for r-dna? The investigator/ lab contact EHSO-Research Safety Institutional Biosafety Committee

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12 6. Components of the r-dna Cheat Sheet Cites the Section from the NIH Guidelines What type of review is required Citation in the Notice of Intent (includes verbiage from the NIH Guidelines) Examples were benchmarked from multiple academic sources, the NIH Guidelines document, and the OBA website

13 Section III A of the NIH Guideline Section Review Required III-A IBC Approval, RAC Review, NIH Director Approval Question on NOI Form / Mirrors NIH Guidelines (10.2) Do any rdna experiments involve the deliberate transfer of a drug resistance trait to microorganisms that are not known to acquire the trait naturally (if such acquisition could compromise ability to control disease agents in humans, veterinary medicine, or agriculture)? Explanation and Examples These experiments are considered Major Actions. Includes: Modifying Mycobacterium tuberculosis so that it can no longer be treated with the Rifampin. Cloning a gene for Erythromycin resistance into Borrelia burgdofieri; Cloning a gene for Chloramphenicol resistance into Rickettsia typhi; Cloning a gene for Pyrimethamine resistance into Toxoplasma gondi; Does not include: Adding an ampicillin resistance gene into a plasmid for selection purposes.

14 Section III B of the NIH Guideline SectionReview Required III-A III-B-1 IBC OBA and Approval, IBC RAC Approval Review, NIH Director Approval Question on NOI Form Explanation and Examples (10.2) (10.3) Do any rdna experiments involve the the These Includes: experiments Cloning experiments are considered for Major the biosynthesis Actions. of deliberate cloning of transfer toxin molecules of a drug resistance with LD50 trait to of Includes: toxin molecules lethal for vertebrates. microorganisms less than 100 ng/kg that are body not weight? known to acquire Modifying Examples: Mycobacterium tuberculosis so that it can no longer the trait naturally (if such acquisition could be -Botulinum treated with toxin the D Rifampin. (LD50=0.4 ng/kg), compromise ability to control disease agents in Cloning -Staphylococcal a gene for enterotoxin Erythromycin B resistance humans, veterinary medicine, or agriculture)? into -Ricin Borrelia burgdofieri; Cloning -Tetrodotoxin a gene for Chloramphenicol resistance into Rickettsia -Tetanus toxin typhi; Cloning -Diphtheria a gene toxin for Pyrimethamine resistance into Toxoplasma -Shigella dysenteriae gondi; neurotoxin -Pertussis toxin Does Specific not approval include: Adding has been an ampicillin given for resistance the cloning gene into a plasmid in Escherichia for selection coli K-12 purposes. of DNA containing genes coding for the biosynthesis of toxic molecules which are lethal to vertebrates at 100 nanograms to 100 micrograms per kilogram body weight.

15 Section III C of the NIH Guideline SectionReview Question on NOI Form Explanation and Examples Required III-A III-C-1 IBC & IRB (10.2) (10.5) Do any rdna experiments involve the the These -Exposing experiments humans are to recombinant considered Major Actions. Approval, deliberate transfer of of a rdna, drug resistance into one trait or to Includes: vaccines, lentiviral vectors (HIV vaccines), adenoassociated Mycobacterium viral vectors, tuberculosis other retroviral so that vectors it can no longer RAC NIH/ORDA Review, microorganisms more human research that are participants? not known to acquire Modifying NIH Registration Director the trait naturally (if such acquisition could -Deliberate treated with transfer the Rifampin. of cell lines to humans Approval compromise ability to control disease agents in Cloning -Introduction a gene of for Markers Erythromycin a cell resistance to compensate for humans, veterinary medicine, or agriculture)? into defective Borrelia genes, burgdofieri; to produce a potentially therapeutic Cloning substance, a gene trigger for Chloramphenicol the immune system resistance to fight into disease Rickettsia -Liposome typhi; use and other methods of delivery Cloning -Synthetic a gene nucleic for Pyrimethamine acid molecules resistance that: into Toxoplasma 1. Contain gondi; more than 100 nucleotides, or 2. Possess biological properties that enable integration Does into not include: the genome, Adding e.g. an cis ampicillin elements resistance involved gene in into a plasmid integration, for selection or purposes. 3. Have potential to replicate in a cell, or 4. Can be translated or transcribed -Delivery of shrna in a plasmid

16 Section III D of the NIH Guideline SectionReview Required III-A III-D-1 III-D-2 III-D-3 Question on NOI Form Explanation and Examples IBC (10.2) (10.6) Do Using any risk rdna group experiments 2, 3, 4 or involve restricted the These NIH experiments Guideline are provides considered risk group Major classification. Actions. Approval, deliberate agents as transfer host-vector of a drug systems? resistance trait to Includes: RAC Review, microorganisms that are not known to acquire Modifying Mycobacterium tuberculosis so that it can no longer NIH Director the (10.7) trait Exposing naturally (if any such animal acquisition to rdna could be Does treated not include: with the Rifampin. exposing mice to human cells that have modified microbes? been transfected with a lentiviral vector previously. Approval compromise ability to control disease agents in Cloning a gene for Erythromycin resistance humans, (10.8) DNA veterinary from Risk medicine, Group or 2, agriculture)? 3, 4 or restricted agents is cloned into a nonpathogenic prokaryotic or lower eukaryotic host vector system? (10.9) The use of infectious DNA or RNA into Cloning Borrelia DNA burgdofieri; from Mycobacterium tuberculosis into a Cloning plasmid a and gene expressing for Chloramphenicol the gene in resistance E. coli (non-k12 into Rickettsia derived strain). typhi; Cloning a gene for Pyrimethamine resistance into Toxoplasma gondi; A helper virus is a virus used to aid in the replication of a viruses or defective DNA or RNA viruses in viral vector. the presence of helper virus in tissue Does not include: Adding an ampicillin resistance gene into a culture systems? plasmid for selection purposes. III-D-4 III-D-5 III-D-6 III-D-7 (10.10) The creation of, (10.11) breeding of, (10.12) experiments with, (10.13) purchase or transfer of transgenic rodents, or other small animals (10.14) Plant research (10.15) Volume Includes: Creating transgenic fruit flies, monkeys, zebrafish, pigs, mosquitos Exotic infectious agents when recombinant or synthetic nucleic acid molecule techniques are associated with whole plants; cloned genomes of exotic agents; clones toxins; involves pathogens of insects (10.16) The use of influenza viruses? Any rdna work with flu

17 Section III D of the NIH Guideline SectionReview Required III-A III-D-1 III-D-2 III-D-3 Question on NOI Form Explanation and Examples IBC (10.2) (10.6) Do Using any risk rdna group experiments 2, 3, 4 or involve restricted the These NIH experiments Guideline are provides considered risk group Major classification. Actions. Approval, deliberate agents as transfer host-vector of a drug systems? resistance trait to Includes: RAC Review, microorganisms that are not known to acquire Modifying Mycobacterium tuberculosis so that it can no longer NIH Director the (10.7) trait Exposing naturally (if any such animal acquisition to rdna could be Does treated not include: with the Rifampin. exposing mice to human cells that have modified microbes? been transfected with a lentiviral vector previously. Approval compromise ability to control disease agents in Cloning a gene for Erythromycin resistance humans, (10.8) DNA veterinary from Risk medicine, Group or 2, agriculture)? 3, 4 or restricted agents is cloned into a nonpathogenic prokaryotic or lower eukaryotic host vector system? (10.9) The use of infectious DNA or RNA into Cloning Borrelia DNA burgdofieri; from Mycobacterium tuberculosis into a Cloning plasmid a and gene expressing for Chloramphenicol the gene in resistance E. coli (non-k12 into Rickettsia derived strain). typhi; Cloning a gene for Pyrimethamine resistance into Toxoplasma gondi; A helper virus is a virus used to aid in the replication of a viruses or defective DNA or RNA viruses in viral vector. the presence of helper virus in tissue Does not include: Adding an ampicillin resistance gene into a culture systems? plasmid for selection purposes. III-D-4 III-D-5 III-D-6 III-D-7 (10.10) The creation of, (10.11) breeding of, (10.12) experiments with, (10.13) purchase or transfer of transgenic rodents, or other small animals (10.14) Plant research (10.15) Volume Includes: Creating transgenic fruit flies, monkeys, zebrafish, pigs, mosquitos Exotic infectious agents when recombinant or synthetic nucleic acid molecule techniques are associated with whole plants; cloned genomes of exotic agents; clones toxins; involves pathogens of insects (10.16) The use of influenza viruses? Any rdna work with flu

18 Section III D of the NIH Guideline SectionReview Required III-A III-D-1 III-D-2 III-D-3 Question on NOI Form Explanation and Examples IBC (10.2) (10.6) Do Using any risk rdna group experiments 2, 3, 4 or involve restricted the These NIH experiments Guideline are provides considered risk group Major classification. Actions. Approval, deliberate agents as transfer host-vector of a drug systems? resistance trait to Includes: RAC Review, microorganisms that are not known to acquire Modifying Mycobacterium tuberculosis so that it can no longer NIH Director the (10.7) trait Exposing naturally (if any such animal acquisition to rdna could be Does treated not include: with the Rifampin. exposing mice to human cells that have modified microbes? been transfected with a lentiviral vector previously. Approval compromise ability to control disease agents in Cloning a gene for Erythromycin resistance humans, (10.8) DNA veterinary from Risk medicine, Group or 2, agriculture)? 3, 4 or restricted agents is cloned into a nonpathogenic prokaryotic or lower eukaryotic host vector system? (10.9) The use of infectious DNA or RNA into Cloning Borrelia DNA burgdofieri; from Mycobacterium tuberculosis into a Cloning plasmid a and gene expressing for Chloramphenicol the gene in resistance E. coli (non-k12 into Rickettsia derived strain). typhi; Cloning a gene for Pyrimethamine resistance into Toxoplasma gondi; A helper virus is a virus used to aid in the replication of a viruses or defective DNA or RNA viruses in viral vector. the presence of helper virus in tissue Does not include: Adding an ampicillin resistance gene into a culture systems? plasmid for selection purposes. III-D-4 III-D-5 III-D-6 III-D-7 (10.10) The creation of, (10.11) breeding of, (10.12) experiments with, (10.13) purchase or transfer of transgenic rodents, or other small animals (10.14) Plant research (10.15) Volume Includes: Creating transgenic fruit flies, monkeys, zebrafish, pigs, mosquitos Exotic infectious agents when recombinant or synthetic nucleic acid molecule techniques are associated with whole plants; cloned genomes of exotic agents; clones toxins; involves pathogens of insects (10.16) The use of influenza viruses? Any rdna work with flu

19 Section III D of the NIH Guideline SectionReview Required III-A III-D-1 III-D-2 III-D-3 Question on NOI Form Explanation and Examples IBC (10.2) (10.6) Do Using any risk rdna group experiments 2, 3, 4 or involve restricted the These NIH experiments Guideline are provides considered risk group Major classification. Actions. Approval, deliberate agents as transfer host-vector of a drug systems? resistance trait to Includes: RAC Review, microorganisms that are not known to acquire Modifying Mycobacterium tuberculosis so that it can no longer NIH Director the (10.7) trait Exposing naturally (if any such animal acquisition to rdna could be Does treated not include: with the Rifampin. exposing mice to human cells that have modified microbes? been transfected with a lentiviral vector previously. Approval compromise ability to control disease agents in Cloning a gene for Erythromycin resistance humans, (10.8) DNA veterinary from Risk medicine, Group or 2, agriculture)? 3, 4 or restricted agents is cloned into a nonpathogenic prokaryotic or lower eukaryotic host vector system? (10.9) The use of infectious DNA or RNA into Cloning Borrelia DNA burgdofieri; from Mycobacterium tuberculosis into a Cloning plasmid a and gene expressing for Chloramphenicol the gene in resistance E. coli (non-k12 into Rickettsia derived strain). typhi; Cloning a gene for Pyrimethamine resistance into Toxoplasma gondi; A helper virus is a virus used to aid in the replication of a viruses or defective DNA or RNA viruses in viral vector. the presence of helper virus in tissue Does not include: Adding an ampicillin resistance gene into a culture systems? plasmid for selection purposes. III-D-4 III-D-5 III-D-6 III-D-7 (10.10) The creation of, (10.11) breeding of, (10.12) experiments with, (10.13) purchase or transfer of transgenic rodents, or other small animals (10.14) Plant research (10.15) Volume Includes: Creating transgenic fruit flies, monkeys, zebrafish, pigs, mosquitos Exotic infectious agents when recombinant or synthetic nucleic acid molecule techniques are associated with whole plants; cloned genomes of exotic agents; clones toxins; involves pathogens of insects (10.16) The use of influenza viruses? Any rdna work with flu

20 Section III D of the NIH Guideline SectionReview Required III-A III-D-1 III-D-2 III-D-3 Question on NOI Form Explanation and Examples IBC (10.2) (10.6) Do Using any risk rdna group experiments 2, 3, 4 or involve restricted the These NIH experiments Guideline are provides considered risk group Major classification. Actions. Approval, deliberate agents as transfer host-vector of a drug systems? resistance trait to Includes: RAC Review, microorganisms that are not known to acquire Modifying Mycobacterium tuberculosis so that it can no longer NIH Director the (10.7) trait Exposing naturally (if any such animal acquisition to rdna could be Does treated not include: with the Rifampin. exposing mice to human cells that have modified microbes? been transfected with a lentiviral vector previously Approval compromise ability to control disease agents in Cloning a gene for Erythromycin resistance humans, (10.8) DNA veterinary from Risk medicine, Group or 2, agriculture)? 3, 4 or restricted agents is cloned into a nonpathogenic prokaryotic or lower eukaryotic host vector system? (10.9) The use of infectious DNA or RNA into Cloning Borrelia DNA burgdofieri; from Mycobacterium tuberculosis into a Cloning plasmid a and gene expressing for Chloramphenicol the gene in resistance E. coli (non-k12 into Rickettsia derived strain) typhi; Cloning a gene for Pyrimethamine resistance into Toxoplasma gondi; A helper virus is a virus used to aid in the replication of a viruses or defective DNA or RNA viruses in viral vector the presence of helper virus in tissue Does not include: Adding an ampicillin resistance gene into a culture systems? plasmid for selection purposes. III-D-4 III-D-5 III-D-6 III-D-7 (10.10) The creation of, (10.11) breeding of, (10.12) experiments with, (10.13) purchase or transfer of transgenic rodents, or other small animals (10.14) Plant research (10.15) Volume (10.16) The use of influenza viruses? Includes: Creating transgenic fruit flies, monkeys, zebrafish, pigs, mosquitos Exotic infectious agents, synthetic nucleic acid molecule techniques, cloned genomes of exotic agents, cloned toxins, pathogens of insects Any rdna work with flu

21 Special cases (many of them!) Appendix B-V. Animal Viral Etiologic Agents in Common Use Murine leukemia virus Not associated with disease in healthy adult humans; Commonly used in laboratory experimental work.

22 Special cases (many of them!) Section III D-4 Caenorhabditis elegans Free-living, non-parasitic soil nematode Commonly used in laboratory experimental work to generate mutants Lamprey

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24 Section III E of the NIH Guideline SectionReview Required Question on NOI Form Explanation and Examples III-A III-E IBC Notice (10.2) Do 10.37: any rdna experiments involve the These All other experiments research are considered Major Actions. Approval, (10.33) deliberate Not transfer fall under of a Sections drug resistance III-A, B, trait C, to D, Includes: Experiments in which the components are F RAC Review, microorganisms that are not known to acquire Modifying nonpathogenic Mycobacterium prokaryotes tuberculosis and non-pathogenic so that it can longer NIH Director the trait naturally (if such acquisition could be lower treated eukaryotes with the Rifampin. Approval (10.34) compromise Involve ability the to formation control disease of rdna agents in Cloning a gene for Erythromycin resistance molecules humans, veterinary containing medicine, no more agriculture)? than 2/3 of into Borrelia burgdofieri; the genome of any eukaryotic virus Cloning a gene for Chloramphenicol resistance into Rickettsia typhi; (10.35) Involve whole plants Cloning a gene for Pyrimethamine resistance into Toxoplasma gondi; (10.36) Involve the creation of transgenic or knock out rodents in which the animal s genome has been altered by stable introduction of rdna or derived from, into the germ-line (10.37) Involve breeding of rodents from 2 strains to generate a new strain or knockout that can be housed at ABSL1 and does not fall under the exempt Appendix C-VIII Does not include: Adding an ampicillin resistance gene into a plasmid for selection purposes.

25 7 Plans for roll-out Tested with investigators Obtain feedback from Investigators and IBC Revise accordingly 8 Ongoing Process Feedback from investigators New technology New Questions from Investigators Revision on NIH Guidelines

26 Other resources available to investigators when classifying their research under the NIH Guidelines Emory University Institutional Review Guidelines IACUC, IRB, Chemical Safety, Radiation Safety Use of viral Vectors in vivo Biosafety Officer NIH OBA website FAQs NIH Guidelines and the use of animals (Very Useful!)

27 Questions Contact Information Esmeralda Meyer & Kalpana Rengarajan Biosafety Office Emory University