Assisting Your IBC in the Assessment and Review of Biosafety Levels in Research with Recombinant DNA

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Assisting Your IBC in the Assessment and Review
of Biosafety Levels in Research with Recombinant
DNA

• Discussion Leaders
• Philip G. Hauck, Mount Sinai School of Medicine
• LouAnn C. Burnett, Vanderbilt University

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Suggested Outline for Discussion

• Finding and registering recombinant DNA research
  at your entity.
• Conducting a review.
• OPTIONAL Case Studies

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Finding rDNA research and getting it registered
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Institutional Resources

• Grants Contracts office
• Animal Care Committee
• Human Subjects Protection Committee
• Other committees (radiation safety, etc.)
• Departmental administrators (chairs, business
  managers, etc.)

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Other Institutional Resources Suggested During
Breakout Session

• Occupational Medicine
• Faculty lists
• NIH Database
• Clinical Trial Coordinators
• USDA Reports
• Purchasing
• Facilities Department
• Vendors (e.g., biosafety cabinet certifiers)
• Research Safety Officer

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Communication with Investigators

• What works? (from discussion)
• Orientation
• One-on-One (as opposed to mass trainings)
• New PIs
• Every new employee gets letter from EHS director
• Group protocol meetings
• Lab-by-lab training
• Walk-throughs
• Hazard checklist

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Reviewing Research according to the NIH Guidelines
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FIRST. . .

• Before you look at the Guidelines, figure out
  what materials are being used and how that
  affects risk.
• THEN, apply the Guidelines

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Risk Groups vs. Biosafety Levels

• Risk Groups
• Organism-specific
• Assigned without regard to how organism will be
  used.
• Biosafety Level
• The combination of laboratory practices, safety
  equipment and laboratory facilities that will
  appropriately contain the organism as it is being
  manipulated.

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Risk Assessment

• How does the Risk Group assignment change when
  other things change (genetic manipulation,
  attenuation, introduction into host organism,
  etc.)?
• What other factors influence the risk of the
  experiment?
• What containment level is needed to minimize
  these risks?

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Containment Goals

• Minimize risks to personnel, etc.
• Contain genetic material.

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Keys to rDNA Review

• Components of the construct
• Vector
• Gene(s)
• Use of the construct (where does it go?)
• Hosts
• Microbe, cell, animal, plant, human?
• Other intentional releases
• Potential for unintentional release

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Reviewing the Construct - Vector

• Vector
• Backbone
• Origin/source
• Promoter (does it change host range?)
• Antibiotic resistance
• Attenuation (is it adequate?)

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Reviewing the Construct - Gene

• Gene(s)
• Source
• Function known/unknown
• Toxin
• Oncogene
• From Risk Group 2, 3, or 4 agent
• Drug resistance trait

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Vector Gene
Biological Containment
Primary Barrier(s)
Secondary Barrier(s)
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Where does the construct go?

• Hosts (biological containment)
• Bacteria
• Conjugative/natural exchanger
• Cells other risks (human)
• Animals
• Plants
• Humans human gene transfer

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Other Considerations

• Human materials
• Select Agents High Consequence Livestock and
  Plant Pathogens
• Animal pathogens

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Other red flags or points of concern (generated
during breakout session)

• Multiples or variations of genes (libraries)
• Transposable elements

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Need more data (generated during breakout
session)

• Competency evaluations
• Practical exams?
• Information on shedding

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Other Intentional Releases

• Field releases, etc.

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Unintentional Releases

• What happens if. . .
• A researcher has a needlestick or other injury
  involving a recombinant DNA construct?
• The construct is released into the environment?
• Are there adequate procedures to address these
  possibilities?

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Who is using it?

• Experienced, TRAINED personnel?
• Vulnerable population?
• How is the experience and/or vulnerability of the
  user assessed? Should others be involved in the
  protocol?

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What about exemptions?

• Dont worry about exemptions
• Protocols are still Biosafety Level 1 (at least),
  even if exempt.
• The BSO (at least) should know what the
  laboratory is working with.

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Each review is case-by-case. . .

• There are very few instances where a rDNA review
  is the same as another rDNA review.
• Each of these key steps must be examined and
  addressed.
• With experience and attention, this can be done
  quickly.
• However. . .technology changes quickly dont
  get complacent!

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Resources

• NIH Guidelines (of course!)
• Biosafety in Microbiological Biomedical
  Laboratories (BMBL)
• www.absa.org
• In development please share your ideas
• Resources Tools/Key Topics Links/IBC
• Resources Tools/Training Tools
• BIOSAFTY email discussion group
• Resources Tools/Email Groups

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Other Resources?
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Examples to share?
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What are the red flags?

• Plasmid expression system with genes that clone
  for Clostridium perfringens epsilon toxin
• Replication-deficient lentiviral vector (HIV-1)
  using helper plasmids
• MoMLV retrovirus with CMV promoter
• Other examples?