Complying with NIH Guidelines at Calvin

1. Complying with NIH Guidelines at Calvin Research & Teaching

2. What this training covers • Various sections of the NIH Guidelines including: • NIH Overview • Calvin’s Institutional Biosafety Committee (IBC) • Recombinant DNA (rDNA) & Synthetic Nucleic Acid Molecule guidelines • Animal guidelines • Plant guidelines • Roles & Responsibilities • Resources • For the purposes of this training, Principal Investigator (PI) includes any faculty or researchers conducting research or teaching coursework at the college

3. NIH Guidelines for Recombinant DNA (rDNA) & Synthetic Nucleic Acid Molecules (SNA)r/sNA • Details procedures and practices for the containment and safe conduct of various forms of rDNA & SNA research. • First drafted in 1976 as an outcome of a meeting of scientists concerned about addressing the potential public health and environmental risks associated with rDNA. • Frequently amended to reflect evolving scientific understanding of rDNA and its applications.

4. The NIH Office of Biotechnology Activities (OBA) • A core responsibility of OBA is to promote awareness of, and adherence to, the standards and practices set forth in the NIH Guidelines. • Provide guidance to IBCs in the oversight of r/sNAresearch • Disseminate information on technical and policy matters concerning r/sNA research • Develop and contribute to public policy on r/sNA research

5. Training Requirements • According to the NIH Guidelines • “The institution is responsible for ensuring that the Principal Investigator (PI) has sufficient training” regarding laboratory safety and implementation of the NIH Guidelines. • “The PI is responsible for ensuring that laboratory staff are appropriately trained.” • We have developed this online training course to fulfill the training requirements for PIs; this will be tracked by the Biosafety Officer (BSO, Lori Keen) in the IBC database. • PIs will be responsible for training their staff and will have access to a downloadable ppt.

6. Course Objectives • Upon completion of this course, participants will have a better understanding of: • The IBC review and approval processes that apply to various forms of research and teaching • The biosafety levels of the NIH Guidelines • The various types of research & teaching covered by the NIH Guidelines • The roles and responsibilities of investigators under the NIH Guidelines

7. The IBC at Calvin College Institutional Biosafety Committee Training

8. The Institutional Biosafety Committee (IBC) • IBCs were established under the NIH Guidelines to provide local review and oversight of research using r/sNA. • They ensure that r/sNAresearch conducted at or sponsored by the institution is in compliance with the NIH Guidelines. • Many institutions, including Calvin, have chosen to assign their IBCs the responsibility of reviewing other research & teaching involving other biohazardous risks (e.g. infectious agents and human samples).

9. Calvin’s IBC • Made up of faculty, staff and community members • Members include representatives from: • Biology and/or Biochemistry- Dave Benson, Curt Blankespoor, John Ubels • Biology Lab Manager - Lori Keen • Environmental Health & Safety – Heather Chapman • 2 community members – David LaGrand, Bob Leunk

10. Calvin College IBC Staff • IBC Coordinator, Lori Keen • Liaison between the investigators & the IBC • Assists faculty with the protocol review process • Notifies investigators of IBC review outcome • Ensures committee operations comply with federal and state regulations

11. IBC Notification Process PIs must notify the IBC prior to commencing your research or teaching, if you plan to use: • Recombinant DNA or Synthetic Nucleic Acid molecules • Infectious agents (BSL2) • Human body fluids or tissues

12. IBC Approval Process • If your research or teaching is non-exempt, you must complete the Biosafety Application • If your research or teaching is exempt, you must complete the IBC First Assessment Form (keep in mind that you may have already completed this) • See next slide for definitions of exempt vs non-exempt • Both forms are to be submitted to the IBC Coordinator, Lori Keen who will ensure the appropriate review process

13. Exempt vs. Non-Exempt Per Section III-F of the NIH Guidelines, experiments are exempt when they involve recombinant DNA or SNA that: • Is not in organisms or viruses; • Uses a host vector system such as: E. coli K 12, Saccharomyces cerevisiae, Saccharomyces uvarum or Bacillus subtilis; and their plasmids; • Uses rDNA molecules containing less than one-half of any eukaryotic genome that are propagated and maintained in cells in tissue culture; • Consist entirely of DNA segments from a single nonchromosomal or viral DNA source; though one or more of the segments may be a synthetic equivalent; • Consist entirely of DNA from a prokaryotic host including its indigenous plasmids or viruses when propagated only in that host (or a closely related strain of the same species); or when transferred to another host by well-established physiological means; • Consist entirely of DNA from a eukaryotic host including its chloroplasts, mitochondria, or plasmids (but excluding viruses) when propagated only in that host (or a closely related strain of the same species). • Details on certain other experiments that may be exempt, as well as exceptions, can be found in Appendix C of the NIH Guidelines (http://oba.od.nih.gov/oba/rac/Guidelines/APPENDIX_C.htm)

14. Exempt vs. Non-Exempt Per Section III of the NIH Guidelines , research or teaching that meet any of the criteria below must submit a proposal to the IBC and receive IBC approval: • Experiments involving the introduction of rDNA or SNA into risk group 2 organisms (includes adenovirus & murine retroviral vectors) or higher • Experiments in which DNA from human or animal pathogens (risk group 2 or higher) is introduced into a non-pathogenic host vector system • Experiments involving the use of infectious DNA or RNA viruses or defective DNA or RNA viruses (risk group 2 or higher) in the presence of helper virus in tissue culture systems • Experiments involving formation of rDNA molecules containing no more than 2/3 of the genome of only eukaryotic virus (with some restrictions). It must be demonstrated that cells lack helper virus • The deliberate transfer of a drug resistance trait to microorganisms that are not known to acquire the trait naturally, if such acquisition could compromise the use of the drug to control disease agents in humans, veterinary medicine or agriculture. • Cloning of toxic molecules with LD50 of less than 100 nanograms/kg body weight • Experiments involving cultures of more than 10 liters • Whole animals in which the animal’s genome has been altered by the stable introduction of rDNA, or RNA derived therefrom, into the germline (transgenic animals) • Experiments involving whole plants • Gene transfer experiments in humans • For more information, you may view Section III at: http://oba.od.nih.gov/oba/rac/Guidelines/NIH_Guidelines.htm

15. IBC Approval Process • The Biosafety Officer can review and approve research or teaching involving: • BSL1 agents, <10 L • Human blood and/or tissue (requires consult with the BSO) • Already approved infectious agents • Environmental swabbing for culturing • If any of the following criteria are met, the research or teaching must be reviewed by the full IBC at a convened meeting: • BSL 2 or higher agents • > 10 L of an agent • Recombinant DNA that is NOT exempt

16. IBC Approval Process • Four possible outcomes: • Approved as is • Approved with conditions • Minor corrections required • Tabled • More information required • Denied • Major corrections and/or information needed. PI generally instructed to consult BSO and re-submit application.

17. IBC Submission • Due to the low volume of Non-exempt work at the College, the IBC meets as necessary, but at least annually to review protocols and annual reports. • If you submit an application, the IBC will convene within 2 weeks to review your application • Applications that do not require full committee review may be submitted to the IBC Coordinator or the IBC Chairperson at any time.

18. IBC Renewal Process • PIs must submit annual reports for projects falling within the NIH Guidelines • The PI will receive a reminder email at least 30 days prior to the report deadline • Approved non-exempt long-term projects must be renewed every 3 years

19. IBC Renewal Process • The BSO can review and approve: • Closures • Renewals without change • Renewals with minor change • Such as personnel changes or changing project titles • The IBC reviews and approves: • Renewals with major changes • Such as an adding an agent to a project, going from in vitro to in vivo • 3-year renewals

20. The NIH Guidelines for Research Involving Recombinant DNA & Synthetic Nucleic Acid Molecules

21. Contents of the NIH Guidelines • Section I –Scope of the Guidelines • Section II –Safety Considerations for r/sNA research • Section III –Types of Experiments Covered by the NIH Guidelines and the levels of review these experiments require • Section IV –Roles and Responsibilities of individuals and entities involved in the conduct and oversight of r/sNA research • Appendices –large number of appendices covering particular topics in greater detail

22. Purpose of the Guidelines • To specify the practices for constructing and handling: • (i) recombinant nucleic acid molecules, • (ii) synthetic nucleic acid molecules, including those that are chemically or otherwise modified but can base pair with naturally occurring nucleic acid molecules, and • (iii) cells, organisms, and viruses containing such molecules

23. Section I - Scope • 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; • (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.

24. Section I –Scope Compliance with the NIH Guidelines • All institutions that receive NIH funding for r/sD research must comply with the NIH Guidelines. • All research & teaching at an institution that are subject to the NIH Guidelines must comply with the requirements even if their individual projects are not funded by NIH. • Compliance with the NIH Guidelinesis mandatory as a condition of receiving NIH funding. • “Guidelines” does not mean “optional”

25. NIH Guidelines – Section II • Safety Considerations • Risk assessments: (Appendix B) - Appendix B lists human etiologic agents and zoonotic agents based on RG RG 1 RG 2 RG 3 RG 4

26. Section II –Safety Considerations • Physical Containment • Defined by 4 Biosafety Levels • BSL1, the least stringent; BSL4, the most stringent • These biosafety levels consist of a combination of • Lab practices and techniques, • Safety equipment, and • Lab facilities appropriate for the operations being performed • Consistent with the CDC Biosafety in Microbiological and Biomedical Laboratories • Described in detail in Appendix G

27. Summary of Recommended Biosafety Levels for Infectious Agents

28. Select Agents • Select Agents are pathogens or toxins considered to have potential as bioweapons. They are regulated by CDC and/or USDA and require approval from those organizations to possess, use, receive or ship them. • A list of Select Agents, and information on the Select Agent Program is available at http://www.selectagents.gov/

29. Section II –Safety Considerations • Biological Containment • Refers to the application of highly specific biological barriers (appendix I) • Limit infectivity of a vector for specific hosts • Limits dissemination and survival in the environment • Example – genetically designed to be replication defective outside of host

30. Risk Group and Biosafety Level are NOT the same • Risk Group (RG) is a stable comparative descriptor of the inherent pathogenic nature of a given microorganism. RG designation is based on how a particular organism is associated with disease in a “healthy” adult and whether prevention or intervention exists for that organism. RG does not change based on how or where the agent is used. • Biosafety Level (BSL) is a variable comparative descriptor of the facility, equipment, practices that serve to “contain” a microorganism, and to ensure the safe use of that organism, while it is being handled. BSL is based on risk assessment and technical judgment and may vary with the use of the agent.

31. Section III –Experiments Covered • Six categories of experiments involving r/sNA based on the level of review required (III-A, III-B, III-C, III-D, III-E, III-F) • Experiments which pose great risk to human health require the highest level of review (category III-A), such as the deliberate transfer of antibiotic resistance into a pathogen, that would then compromise the use of that drug to treat the disease. • Experiments that are not considered to pose a risk to human health or the environment are exempt from the Guidelines(category III-F) and do not require review (e.g., the purchase of transgenic animals that can be housed at BSL1).

32. Six Levels of Review

33. Dual Use Research Life sciences research that yields information or technologies with the potential to be misused to threaten public health or national security. • The National Science Advisory Board for Biosecurity (NSABB) has identified seven categories of experiments that have the potential to fall under Dual Use Research of Concern: • Enhance the harmful consequences of a biological agent or toxin. • Disrupt immunity or the effectiveness of an immunization without clinical and/or agricultural justification. • Confer resistance to prophylactic or therapeutic interventions, or facilitate the ability of an agent or toxin to evade detection methodologies. • Increase the stability, transmissibility, or the ability to disseminate a biological agent or toxin. • Alter the host range or tropism of a biological agent or toxin. • Enhance the susceptibility of a host population. • Generate a novel pathogenic agent or toxin or reconstitute an eradicated or extinct biological agent.

34. Recombinant and Synthetic Nucleic Acid Research Involving Animals and the NIH Guidelines

35. Section III-D-4 Experiments Involving Animals • IBC and Institutional Animal Care & Use Committee (IACUC) Approval Before Initiation • Includes Experiments in which: • the animal’s genome has been altered by stable introduction of r/sNAinto germline (i.e., transgenic animals) • applies to both the generation and use of transgenic animals (including arthropods) • r/sNA modified microorganisms are tested on whole animals • BSL2 or BSL2-N or greater containment

36. Section III-E-3 Experiments Involving Transgenic Rodents • IBC and IACUC Approval Before Initiation at Calvin • Includes Experiments in which: • Rodent’s genome has been altered by stable introduction of recombinant or synthetic nucleic acid into germline • applies only to the generation of transgenic rodents • BSL1 containment is appropriate

37. Appendix C-VI Exempt Animal Research • The purchase or transfer of transgenic rodents that require BSL-1 containment • Further manipulations of these animals with r/sNA are not necessarily exempt from the NIH Guidelines

38. IBCs and IACUCs and Animal Research • Joint purview and collaborative review over certain types of research • Transgenic or cloned animals • Use of recombinant or synthetic nucleic acid molecules in animals • Pre-clinical studies and data assessment for human gene transfer protocols • At Calvin, the EHS Officer is a member of both committees

39. IBC and IACUC Review of Animal Research Utilizing r/sNA

40. Points to Consider for Animal Research with r/sNA • Containment procedures (SOPs) • Physical and biological • Plans for recapture of escapees • Consequences should containment fail • Procedures for transfer and transportation of animals • Disposal and destruction methods • Breeding SOPs • Occupational biosafety concerns • Personal protective equipment • Decontamination

41. Appendix Q –Physical and Biological Containment for Large Animals • Applies when research animals are of a size or have growth requirements that preclude laboratory containment • For example, cattle, swine, sheep, goats, horses, poultry, etc. • Addresses containment and confinement practices in animal facilities (BL1-N to BL4-N) • Applies to animals: • In which genome is altered by stable introduction of rDNA or SNA; or • On which rDNA or SNA-modified microorganisms are being tested

42. Recombinant & Synthetic Nucleic Acid Research Involving Plants and the NIH Guidelines

43. Section III-D-5 Experiments Involving Plants • IBC Approval Before Initiation • Includes Experiments in which: • Plants are genetically engineered by r/sNAor synthetic biology methods, or • Plants are used with r/sNA modified insects • Generally BL1-P through BL4-P, depending on risk

44. IBC Approval Before Initiation • Includes Experiments: • Involving r/sNA-modified whole plants • That are not noxious weeds and will not hybridize with noxious weeds, and • For which BL1-P containment is appropriate

45. Appendix P –Physical and Biological Containment for Plants • Defines physical containment levels for r/sNA research involving plants • Describes the four plant Biosafety Levels BL1-P through BL4-P and provides details on the standard practices and any special procedures that should be followed • Describes biological containment practices for r/sNA-containing plants, plant-associated microorganisms, and plant-associated small animals.

46. Section IV of the Guide

47. Section IV –Roles and Responsibilities • Institution • Institutional Biosafety Committee (IBC) • Biological Safety Officer (BSO) • Principal Investigator (PI) • NIH

48. Institutional Responsibilities • Establish and implement policies for the safe conduct of recombinant or synthetic nucleic acid research • Establish an IBC • Ensure compliance with the NIH Guidelines by investigators • Ensure appropriate training for IBC members and staff, PIs, and laboratory staff • Determine necessity for health surveillance of personnel • Report any significant incidents or violations to the NIH Guidelines to OBA within 30 days

49. IBC Responsibilities • Assessment of r/sNAresearch • Conformity with the NIH Guidelines • Potential risk to environment and public health • Containment levels per NIH Guidelines • Adequacy of facilities, SOPs, PI and lab personnel training • Institutional and investigator compliance; e.g., adverse event reports • Periodically review r/sNAuse for compliance • This may include periodic unannounced inspections • Adopt emergency plans covering spills, contamination, other accidents • The IBC may not authorize initiation of r/sNA experiments not explicitly covered by the NIH Guidelines until NIH establishes the containment requirement

50. IBC Membership • Membership requirements • At least 5 individuals • Appropriate r/sNA expertise • Plant and animal experts if needed, biosafety officer as appropriate • At least two members not affiliated with the institution • represent the interest of the surrounding community with respect to health and protection of the environment • Laboratory technical staff (recommended) • Expertise in assessment of risk to environment and public health • Knowledge biological safety and physical containment