Download March 2013

March 2013
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October 2011 NIH rDNA Guidelines (and
numerous rDNA and IBC support documents)
◦ http://www4.od.nih.gov/oba/rdna
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Changes effective in March 2013
◦ Scope expanded to include synthetic nucleic acids
◦ Title of Guidelines will change (…recombinant or
synthetic nucleic acid molecules…)
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National Science Advisory Board for Biosecurity
◦ Dual use, synthetic biology, personnel reliability,
biosecurity of select agents
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Scientifically responsive document
PI is responsible for full compliance with the
NIH Guidelines in the conduct of rDNA
research
Continue to evolve (current edition October
2011, next update March 2013)
Guidelines = term/condition of funding from
NIH!
◦ Consequences of non-compliance can be significant
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May not cover every experiment – call OBA for
Guidance
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Scope and Applicability
◦ Practices for constructing and handling
 Recombinant and Synthetic Nucleic Acid Molecules
 Organisms and viruses containing recombinant and
synthetic nucleic acid molecules
◦ Definition
 Constructed outside living cells by joining natural or
synthetic DNA segments to DNA molecules that can
replicate in a living cell
 Molecules resulting from the replication of those
described above
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Section I-C-1-a-(1). Research that is
conducted at or sponsored by an institution
that receives any support for research
involving recombinant and synthetic nucleic
acid molecules from NIH, including research
performed directly by NIH. An individual who
receives support for research involving
recombinant DNA must be associated with or
sponsored by an institution that assumes the
responsibilities assigned in the NIH
Guidelines.
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Section I-D-1. All NIH-funded projects
involving recombinant DNA techniques must
comply with the NIH Guidelines. Noncompliance may result in: (i) suspension,
limitation, or termination of financial
assistance for the noncompliant NIH-funded
research project and of NIH funds for other
recombinant DNA research at the institution,
or (ii) a requirement for prior NIH approval of
any or all recombinant DNA projects at the
institution.
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Identify if Research is subject to the
Guidelines and Identify the Guideline
Section
Propose an appropriate Containment
Level (BSL)
Obtain Yale Biological Safety Committee
Approval before starting work subject to
the NIH Guidelines
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Train lab staff in safe practices and
emergency response procedures
Alert staff of any special medical
surveillance restrictions or immunizations
Supervise and monitor lab staff for
adherence to safety protocols
Request authorization for significant
updates of your rDNA experiments from
the Yale Biological Safety Committee
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Report any significant problems, such as
violations of the NIH Guidelines or any
significant research related accidents,
exposures and illnesses to:
◦ Yale Biological Safety Committee (203-7853550)
◦ Yale EHS Biosafety Office (203-785-3550)
◦ NIH Office of Biotechnology Activities (301496-9838
Risk Group 1
(RG1)
Risk Group 2
(RG2)
Agents that are not associated with disease in
healthy adult humans
Agents that are associated with human disease
which is rarely serious and for which preventive or
therapeutic interventions are often available
Risk Group 3
(RG3)
Agents that are associated with serious or lethal
human disease for which preventive or
therapeutic interventions may be available (high
individual risk but low community risk)
Risk Group 4
(RG4)
Agents that are likely to cause serious or lethal
human disease for which preventive or therapeutic
interventions are not usually available (high individual
risk and high community risk)
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Classification of parent agent
Toxins
Antibiotic resistance genes
Altered host range or tropism
Replication competency
Integration into host genome
Interference with cell cycle
Toxicity, allergenicity, other
Level of Review
Example of rDNA Experiment
NIH Guideline
Section
IBC, RAC review and NIH
Director Approval
Experiments that compromise the
control of disease agents in medicine
through deliberate transfer or a drug
resistance trait
III-A
IBC Approval and NIH
review
Deliberate formation of rDNA
containing genes of a toxin with an
LD50 < 100 ng/kg
III-B
IBC and IRB Approval and
NIH review
Introduction of rDNA into human
subjects (Human gene transfer)
III-C
IBC approval before
initiation
Wide range (rDNA exp’s involving
pathogens, defective vectors,
animals, plants, large scale
III-D
IBC Notice at initiation
Creating transgenic rodents, low risk
rDNA Plant experiments
III-E
Exempt – registration not
required
Those do not represent a significant
risk to health or the environment
III-F
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III-A-1: Deliberate transfer of a drug
resistance trait that could compromise
disease control
III-B-1: Cloning of toxins with low LD50
(<100 ng/kg)
rDNA experiments involving restricted agents
◦ DNA from restricted agents into host cells
◦ Experiments introducing DNA into restricted agents
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Experiments involving restricted poxviruses
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III-C-1: Human gene transfer experiments
Experiments not explicitly covered by the NIH
rDNA Guidelines (NIH OBA establishes
containment)
Changes in containment level that varies from
what has been specified in the NIH rDNA
Guidelines
Protocols that have been approved at another
institution as a Major Action by the NIH
◦ i.e. Location B cannot approve the same work that
has been approved at Location A by the NIH (w/out
NIH permission) (will change March 2013)
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Section III-D
◦ Use of Risk Group 2 or higher pathogens as
host or vector
◦ Cloning DNA or RNA from Risk Group 2
pathogens
◦ Use of defective pathogen vectors with helper
virus or packaging cells
◦ rDNA experiments involving whole animals
(arthropods)
◦ rDNA experiments involving whole plants
◦ Large Scale (> 10 Liters) rDNA experiments
◦ Experiments involving high risk Influenza
viruses
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Section III-E (Experiments that can be
initiated at the time of registration)
◦ rDNA Experiments that involve the formation of
rDNA molecules that contain less than 2/3’s of any
Eukaryotic Virus
◦ Experiments involving whole plants (not covered
under III-D)
◦ Creation of Transgenic animals
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Section III-F (Exempt rDNA Experiments)
◦ Appendix C
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Vector
◦ Risk Group of parent agent
◦ Defective/Replication incompetent
 Chance to recover missing portions
 Test for replication competency prior to use
◦ Percentage of the genome utilized
◦ Shedding of the vector in animal experiments
◦ Risks associated with exposure to vector
components
◦ Integration into host cells or transient infection
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Inserted DNA
◦ Structural genes
◦ Cell cycle/regulatory genes
◦ Oncogenes
 Human, animal
◦ Toxins
◦ Genomes from other pathogens
 Chapter 28 Cell Lines: Biosafety and Viral Gene
Transfer Vectors (Tom Kost, Patrick Condreay and
Claudia Mickelson)
 ASM Biosafety: Principles and Practices, 4th Edition
Fleming, D.F. and Hunt, D. Editors
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Risk of host cell line starting point
Consider insert?
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Viral sequences
Pathogenic function
Virulence factors
Transactivation of endogenous viruses
Production of highly reactive bioactive molecules
 Growth factors
 Growth factor receptors
 Recombinant gene products
Replication defective vectors
◦ Competent virus may arise in cultures – must
monitor
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Host Cells
◦ Human or Non-human primate
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Primary or continuous
Transformed
Shedding human pathogens
Oncogenic
◦ Other mammalian cells (rodent, dog, pig, etc.)
◦ Non mammalian cells (plant, insect, etc.
 Chapter 11 Cell Lines: Applications and Bioafety
(Otto Doblhoff-Dier and Gwyn Stacey)
 ASM Biosafety: Principles and Practices, 4th Edition
Fleming, D.F. and Hunt, D. Editors
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Yale EHS Office
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Biosafety Officers
The Safety Advisor Assigned to your lab
Back-up Safety Advisor(s)
203-785-3550
Yale EHS website
◦ www.yale.edu/ehs
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NIH Office of Biotechnology Activities
◦ Webpage
 http://oba.od.nih.gov/oba/index.html