Summary of NIH Guidelines for Recombinant DNA & Synthetic Nucleic Acids

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1 Summary of NIH Guidelines for Recombinant DNA & Synthetic Nucleic Acids The National Institutes of Health (NIH) Guidelines for Research Involving Recombinant Synthetic Nucleic Acid Molecules (recombinant or synthetic nucleic acids NIH Guidelines) are published by the Office of Biotechnology Activities (OBA). The NIH Guidelines outline the scope of experiments involving recombinant or synthetic nucleic acid molecules and the required containment levels for each category of experiment for the safe conduct of research. The NIH Guidelines are amended periodically to ensure protection of the workers handling recombinant or synthetic nucleic acid molecules, and also to protect public health and the environment, and that adequate containment is in place. The most recent copy of the NIH Guidelines can also be accessed at the NIH OBA website at: Recombinant and synthetic nucleic acid molecules are defined as: Molecules that are constructed by joining nucleic acid molecules and can replicate in a living cell, i.e. recombinant nucleic acids; 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 Molecules that result from the replication of those described above. The NIH Guidelines apply to research that is conducted at or sponsored by an institution that receives NIH funding for research with recombinant or synthetic nucleic acids or institutions that receive funding for molecular biology experiments. Institutions that receive funding from the NIH must ensure full conformity with the NIH Guidelines for all experiments conducted at the institution regardless of funding source. Failure to comply with the NIH Guidelines can lead to a loss of NIH funding for recombinant DNA Research and could ultimately result in suspension of NIH research funding awarded to the institution until corrective measures have been established. Key responsibilities for the institution receiving NIH funding for research with recombinant or synthetic nucleic acids include: Establish an Institutional Biosafety Committee (IBC); Appoint an IBC Chair to lead the periodic IBC meetings; Designate a Biological Safety Officer (BSO) when high risk or large scale experiments are conducted; Ensure adequate expertise for the review of recombinant or synthetic nucleic acids protocols involving plants, animals and human gene transfer experiments; Periodically inspect laboratories to ensure that the facility, work practices and other requirements conform with the NIH Guidelines; Train IBC members, the BSO, Principal Investigators, and laboratory staff; Develop a health surveillance program for researchers conducting high risk and large scale experiments; Develop emergency response plans for addressing spills, exposures, laboratory acquired infections, containment breaches, and violations; File an annual update of the IBC roster with biosketches to the NIH; and Report significant exposures, accidents, loss of containment or release of recombinant materials to OBA. The Institutional Biosafety Committee

2 The Institutional Biosafety Committee (IBC) should be established and supported by the highest administrator at the institution. The IBC should have oversight for potentially hazardous work and does not have to be limited to experiments involving recombinant or synthetic nucleic acids. IBCs set the policy for the safe conduct of biohazard research and oversee all experiments involving recombinant or synthetic nucleic acids projects conducted at the institution. IBCs must ensure compliance with the NIH Guidelines throughout the entity at NIH funded institutions. All investigators at an institution that receives NIH funding must comply with the NIH Guidelines regardless of funding source. This is an important point for an institution as compliance with the NIH Guidelines extends to each and every one of their laboratories. All work must be conducted in compliance with the NIH Guidelines. Responsibilities of the Institutional Biosafety Committee (IBC) include the requirement to retain a minimum of 5 members, at least 2 of which must be from the community and not affiliated with the institution. Members should be appointed by the institution's senior responsible official and reviewed annually. It is recommended that IBC members have a set term limit, to ensure a rotation of new members onto the committee at periodic intervals. Members may not participate in the review or approval of any of their own protocols or those in which they hold a financial or other interest. Institutions should have a conflict of interest policy to ensure that this policy is followed. The IBC must also have expertise in molecular biology and recombinant or synthetic nucleic acids technology; human, animal and plant pathogens; biosafety, occupational health and medical surveillance; physical and biological containment; and legal guidance where applicable. IBC membership should include varied disciplines, from recombinant or synthetic nucleic acids and molecular biology, microbiology, laboratory safety, engineering, and representation from the laboratory technical staff. IBC members with plant and animal containment expertise must be available if research involves plants, animals or plant and animal pathogens. IBCs should not be limited to its membership in the review of recombinant or synthetic nucleic acids protocols as the institution should have access to other faculty with expertise when needed. Other experts within the institution and even those outside the institution should be contacted for assistance if needed in the review of recombinant or synthetic nucleic acids research in a non-voting capacity. IBCs can also review experiments with infectious agents, toxins and other biohazards, as they are not limited to only recombinant or synthetic nucleic acids experiments. The IBC must review the PIs written risk assessment and site-specific safety procedures. The PI's written risk assessment should include a proposal for physical and biological containment levels in accordance with the NIH guidelines when registering work with the IBC. The PI must also submit a registration document that contains specific information regarding their proposed non-exempt recombinant or synthetic nucleic acids experiment. The IBC must perform an independent risk assessment and is responsible for setting the containment level for the experiment. The IBC has responsibility for ensuring that Principal Investigators have trained personnel. Retraining should be provided periodically. Although not specified, retraining should be frequent enough to ensure that PIs maintain awareness of their responsibilities under the NIH Guidelines. Inspections of proposed work practices and facilities should also be periodically monitored and documented. IBCs should also develop a policy for sanctions in advance and communicate this to the PIs at their institution. Related institutional entities, such as the Dean's Office, Department Chairs, or Institutional Officials, should be involved in the development of a sanction policy. Securing the support of the administration is integral to an effective program. As non-conformity can involve a multitude of factors, each incident should be evaluated on a case by case basis. IBCs should work with their PIs

3 after each incident to prevent a repeat occurrence of the event. Incidents should be documented, filed and reported to required authorities. In addition to the NIH OBA, the IBC should establish formal relationships with related Committees and groups within and outside their institution, such as the Institutional Review Board (IRB) and the Institutional Animal Care and Use Committee (IACUC). Within an institution, the IBC should have cross representation with the IACUC and the IRB. Problems with the NIH Guidelines identified by the BSO are reported to the IBC. The BSO also advises on laboratory security and assists with the development of emergency response plans. To summarize, the official duties of the BSO under the NIH Guidelines are to: Conduct periodic inspections to ensure that laboratory standards are followed; Report to the IBC any significant problems, violations of the NIH Guidelines, and any significant research-related accidents and illnesses; Develop emergency response plans for handling accidental spills and personal contamination; Investigate laboratory accidents involving recombinant or synthetic nucleic acids and assist with remediation and mitigation measures; Provide advice on laboratory security; and Provide technical advice to PIs and the IBC on research safety procedures. Principal Investigators conducting recombinant or synthetic nucleic acids experiments are responsible for full compliance with the NIH Guidelines. They make the initial NIH recombinant or synthetic nucleic acids classification and determination of containment for their experiments at time of registration. PI's are also responsible for instructing and training researchers in laboratory techniques and safe working procedures and for oversight of the safety performance for the researchers they supervise. The PI cannot begin or modify any non-exempt recombinant or synthetic nucleic acids experiments without approval or in some cases registration by the IBC. The PI must also report violations of the NIH Guidelines and serious recombinant or synthetic nucleic acids incidents to the NIH OBA within 30 days. PIs must conduct a formal risk assessment for their studies. The NIH Guidelines provides assistance with risk assessment in Section II of the NIH Guidelines. Detailed information on Risk Assessment was provided in an earlier Module in the Biosafety Series entitled Biohazard Risk Assessment. PIs are also responsible for selecting the appropriate classification within the NIH Guidelines of their proposed experiments. Section III of the NIH Guidelines describes experiments that require registration, review, and approval before initiation. A summary of the NIH Guidelines Section III is provided below. Section III-A-1 (Major Action) Cloning a therapeutic antibiotic resistance gene into a human, animal, or plant pathogen is a major action, if the transfer could compromise the ability to treat or control the disease if the resistance is not acquired naturally (these experiments must be reviewed by the NIH Recombinant DNA Advisory Committee (RAC) and approved by the NIH Director). Section III-B-1 This section is related to the cloning of DNA encoding for a low LD50 toxin or work with vectors that

4 express toxins with a low LD50 (< 100 nanograms per Kilogram body weight). These experiments must be registered with and approved by the NIH RAC. Botulinum and tetanus toxin are examples of toxins that have LD50's below 100 nanograms per Kilogram of body weight. The cloning of toxins with an LD50 < 100 micrograms per Kilogram body weight must also be reviewed and approved by the IBC. This section also includes experiments that have already been approved as a III-A-1-a Major Action under the NIH Guidelines if the exact experiment is proposed at a new institution. This experiments may not be initiated until approved by the IBC and reviewed by the NIH. Section III-C-1 Human gene transfer research experiments: the deliberate transfer or recombinant or synthetic nucleic acids, or DNA or RNA derived from recombinant or synthetic nucleic acids, into one or more human research participants. These experiments include the transfer of DNA with defective viral vectors, such as retroviral, adenoviral and Lentiviral vectors, along with the use of liposomes and other methods of delivery. These experiments require registration with the NIH OBA and also preapproval from the Institutional Review Board and the U.S. Food and Drug Administration prior to initiation. Section III-D Section III-D experiments comprise the bulk of the experiments reviewed by the IBC and include the use of Risk Group 2 through 4 agents and restricted pathogens, or defective pathogens in cell culture, animals, plants, arthropods, and large scale experiments (work with > 10 liters of recombinant or synthetic nucleic acids culture). Examples of III-D Experiments Cloning a gene into a pathogen Use of a pathogen as a vector or expressing a pathogen into a lower prokaryotic or lower eukaryotic cell Using a defective pathogen vector with or without helper virus in cell culture or animal experiments Experiments involving toxins, pathogens, defective vectors, and other genetically materials used in animals, plants or arthropods. Section III-E Section III-E experiments are a category of research that allow the PI to initiate the work at the time of submission of a recombinant or synthetic nucleic acids registration form to the IBC. The protocol must still be reviewed by the IBC and any new information from the review must be provided to the PI following the IBC meeting. Examples of III-E experiments include: Research that involves less than 2/3 of the genome of Risk Group 2 agents; (Experiments involving Risk Group 3 or 4 pathogens or from restricted agents are not included) The creation or production of transgenic rodents requiring BSL1 containment; and Work with whole plants that involve the use of noxious weeds, non-exotic agents, and exotic agents that won't threaten the ecosystem are also included as III-E experiments. Exempt experiments (represent recombinant or synthetic nucleic acids research that does not pose a

5 significant risk and do not require review and registration by the IBC) are described in Section III-F of the NIH Guidelines. Examples of exempt research includes: Those recombinant or synthetic nucleic acids experiments that are not in organisms or viruses; PCR; The use of known exchangers, recombinant or synthetic nucleic acids from a host organism propagated in the same host; and Those recombinant or synthetic nucleic acids experiments that do not pose a significant risk to health or the environment. Appendix C details exempt experiments that do not pose a risk to the public and include research that involves less than 50% of the genome from a Risk Group 2 pathogen, experiments using E. coli, B. subtilis, or S. cerevisiae, and the purchase or transfer of transgenic rodents. The generation of transgenic rodents by breeding to create a new strain shall be EXEMPT from the NIH Guidelines if the following criteria are met. Both parental rodents can be housed under BSL1 containment; AND Neither parental transgenic rodent contains the following genetic modifications: o Incorporation of more than 50% of the genome of an exogenous eukaryotic virus from a single family of viruses; OR o Incorporation of a transgene that is under the control of a gammaretroviral long terminal repeat (LTR); AND o The transgenic rodent that results from this breeding is not expected to contain more than 50% of an exogenous viral genome from a single family of viruses. This exemption DOES NOT pertain to other transgenic animals such as zebrafish, drosophila, rabbits, pigs, etc. It also DOES NOT pertain to transgenic experiments involving plants. IBC Meetings IBC meetings must be conducted in a manner that allows interaction between committee members either in person or through a conference call. Attendance must be taken at IBC meetings to ensure that a meeting quorum has been attained. IBC meetings are encouraged to be open to the public and institutions are urged to post information regarding their meeting schedules. Minutes of IBC meetings must be filed and made available to the NIH OBA and the public upon request. IBC minutes should include the following information: Date and place of the IBC meeting Record that prior minutes were approved Individuals in attendance Opening and closing times of the meeting All major motions Major points that were approved Time of the meeting opening and adjournment Sufficient detail to record key discussion and the rationale for any decisions made (this should include a review of the PIs risk assessment and detail the following): o Agent characteristics o Type of manipulations planned

6 Incident Reports o Source of the inserted DNA sequences o Nature of the inserted DNA sequences o Host(s) and vector(s) to be used o Whether an attempt will be made to obtain expression of a foreign gene and the protein that will be produced o Containment conditions to be implemented o Section of the NIH Guidelines applicable to the proposed work Significant incidents, violations and research-related accidents and illnesses must be reported to the NIH OBA within 30 days or immediately in the event of personal exposures. Spills or accidents in BSL2 laboratories resulting in overt exposure must be immediately reported to NIH OBA and the IBC. Spills or accidents occurring in high containment laboratories resulting in an overt or potential exposure must be immediately reported to NIH OBA, the IBC and the BSO. Although PIs are responsible for reporting incidents, the institution, IBC and BSO may also report the incident to the NIH OBA. Types of Incidents that Must be Reported Spills, or accidents in BSL2 laboratories resulting in overt exposure must be immediately reported to the NIH OBA Spills, or accidents resulting in overt or potential exposure in BSL3 and BSL4 must be immediately reported to the NIH OBA Personal injury involving recombinant or synthetic nucleic acid molecules Illness potentially associated with recombinant or synthetic nucleic acids research Breach of containment associated with recombinant or synthetic nucleic acids molecules (i.e. a spill of BSL2 or higher recombinant or synthetic nucleic acids outside of primary containment) Improperly discarded recombinant or synthetic nucleic acid waste materials Incident reports to NIH OBA should include the nature and consequences of the incident, the cause of the incident, and a detailed report of the response measures taken by the institution. Appendices of the NIH Guidelines The NIH Guidelines also contain appendices to assist IBCs in their review of recombinant or synthetic nucleic acids protocols. Appendix B contains the "Classification of Etiologic Agents on the Basis of Hazard," which lists human pathogens by risk group Appendix G provides containment requirements for cell culture and small animal experiments and outline appropriate work practices, safety equipment, and facility design for each biocontainment level (BSL1 through BSL4) Appendix K provides containment requirements for large scale recombinant or synthetic nucleic acid experiments (> 10 liters of culture) Appendix M details the requirements for human gene transfer experiments.

7 Appendix P provides containment requirements for recombinant or synthetic nucleic acid experiments involving transgenic plants Appendix Q provides containment requirements for recombinant or synthetic nucleic acid experiments involving large animals