CLASSES OF RECOMBINANT DNA ACTIVITY

Section 61-1.20 - Classes of recombinant DNA activity

CLASSES OF RECOMBINANT DNA ACTIVITY

61-1.20 Classes of recombinant DNA activity. This section discusses activities involving recombinant DNA. The activities are divided into four classes corresponding to those set forth in part III of the revised NIH guidelines effective August 27, 1982. The words experiment and experiments are to be read as referring to recombinant DNA activity as defined in article 32-A of the Public Health Law. The classes and their respective requirements are:

III-A. Experiments which require specific NIH and institutional biosafety committee (IBC) approval before initiation of the experiment.

III-B. Experiments which require IBC approval before initiation of the experiment.

III-C. Experiments which require IBC notification at the time of initiation of the experiment.

III-D. Experiments which are exempt from the NIH guidelines and as such may come under section 61-1.3(c) of this Subpart.

If an experiment falls into both class III-A and one of the other classes, the rules pertaining to class III-A must be followed. If an experiment falls into class III-D and into either class III-B or III-C as well, it can be considered as a class III-D activity.

Changes in containment levels from those specified here may not be instituted without the express approval of the Director, NIH.

III-A Experiments that require NIH and institutional biosafety committee (IBC) approval before initiation. Experiments in this category cannot be initiated without specific approval by NIH. The containment conditions for such experiments will be set by NIH at the time of approval. Such experiments also require the approval of the IBC before initiation. Specific experiments already approved in this category and the appropriate containment conditions are listed in appendices D and F of the NIH guidelines. If an experiment is similar to those listed in appendices D and F, NIH Office of Recombinant DNA Activities (ORDA) may determine appropriate containment conditions according to case precedents.

III-A-1. Deliberate formation of recombinant DNAs containing genes for the biosyntheses of toxic molecules lethal for vertebrates at an LD50 of less than 100 nanograms per kilogram body weight (e.g., microbial toxins such as the botulinum toxins, tetanus toxin, diphtheria toxin, Shigella dysenteriae neurotoxin). Specific approval has been given for the cloning in E. coli K-12 of DNAs containing genes coding for the biosynthesis of toxic molecules which are lethal to vertebrates at 100 nanograms to 100 micrograms per kilogram body weight. Containment levels for these experiments are specified in appendix F of the NIH guidelines.

III-A-2. Deliberate release into the environment of any organism containing recombinant DNA.

III-A-3. Deliberate transfer of a drug resistance trait to microorganisms that are not known to acquire it naturally, if such acquisition could compromise the use of the drug to control disease agents in human or veterinary medicine or agriculture.

III-B Experiments that require institutional biosafety committee (lBC) approval before initiation. Investigators performing experiments in this category must submit to their IBC, prior to initiation of the experiments a registration document that contains a description of: (a) the source(s) of DNA; (b) the nature of the inserted DNA sequences; (c) the hosts and vectors to be used; (d) whether a deliberate attempt will be made to obtain expression of a foreign gene, and, if so, what protein will be produced; and (e) the containment conditions specified in the NIH guidelines. This registration document must be dated and signed by the investigator and filed only with the local IBC. The IBC shall review all such proposals prior to initiation of the experiments. Requests for lowering of containment for experiments in this category will be considered by NIH.

III-B-1. Experiments using human or animal pathogens (Class 2, Class 3, Class 4 or Class 5 agents) as host-vector systems.

III-B-1-a. Experiments involving the introduction of recombinant DNA into Class 2 agents can be carried out at P2 containment.

III-B-1-b. Experiments involving the introduction of recombinant DNA into Class 3 agents can be carried out at P3 containment.

III-B-1-c. Experiments involving the introduction of recombinant DNA into Class 4 agents can be carried out at P4 containment.

III-B-1-d. Containment conditions for experiments involving the introduction of recombinant DNA into Class 5 agents will be set on a case-by-case basis following ORDA review. A USDA permit is required for work with Class 5 agents.

III-B-2. Experiments in which DNA from human or animal pathogens (Class 2, Class 3, Class 4 or Class 5) agents is cloned in nonpathogenic prokaryotic or lower eukaryotic host-vector systems.

III-B-2-a. Recombinant DNA experiments in which DNA from Class 2 or Class 3 agents is transferred into nonpathogenic prokaryotes or lower eukaryotes may be performed under P2 containment. Recombinant DNA experiments in which DNA from Class 4 agents is transferred into nonpathogenic prokaryotes or lower eukaryotes can be performed at P2 containment after demonstration that only a totally and irreversibly defective fraction of the agent's genome is present in a given recombinant. In the absence of such a demonstration, P4 containment should be used. Specific lowering of containment to P1 for particular experiments can be approved by the IBC. Many experiments in this category will be exempt from the NIH guidelines (see III-D). Experiments involving the formation recombinant DNAs for certain genes coding for molecules toxic for vertebrates require NIH approval (see III-A-I), or must be carried out under NIH specified conditions as described in appendix F of the NIH guidelines.

III-B-2-b. Containment conditions for experiments in which DNA from Class 5 agents is transferred into nonpathogenic prokaryotes or lower eukaryotes will be determined by ORDA following a case-by-case review. A USDA permit is required for work with Class 5 agents.

III-B-3. Experiments involving the use of infectious animal or plant viruses or defective animal or plant viruses in the presence of helper virus in tissue culture systems.

Caution:Special care should be used in the evaluation of containment levels for experiments which are likely to either enhance the pathogenicity (e.g., insertion of a host oncogene) or to extend the host range (e.g., introduction of novel control elements) of viral vectors under conditions which permit a productive infection. In such cases, serious consideration should be given to raising the physical containment by at least one level. Note:Recombinant DNA molecules which contain less than two thirds of the genome of any eukaryotic virus (all virus from a single family being considered identical) may be considered defective and can be used, in the absence of helper, under the conditions specified in III-C.

III-B-3-a. Experiments involving the use of infectious Class 2 animal viruses, or defective Class 2 animal viruses in the presence of helper virus, can be carried out at P2 containment.

III-B-3-b. Experiments involving the use of infectious Class 3 animal viruses, or defective Class 3 animal viruses in the presence of helper virus, can be performed at P3 containment.

III-B-3-c. Experiments involving the use of infectious Class 4 viruses, or defective Class 4 viruses in the presence of helper virus, may be carried out under P4 containment.

III-B-3-d. Experiments involving the use of infectious Class 5 viruses, or defective Class 5 viruses in the presence of helper virus, will be determined on a case-by-case basis following ORDA review. A USDA permit is required for work with Class 5 pathogens.

III-B-3-e. Experiments involving the use of infectious animal or plant viruses, or defective animal or plant viruses in the presence of helper virus, not covered by III-B-3-a, III-B-3-b, III-B-3-c or III-B-3-d may be carried out under P1 containment.

III-B-4. Recombinant DNA experiments involving whole animals or plants.

III-B-4-a. DNA from any source except for greater than two thirds of a eukaryotic viral genome may be transferred to any nonhuman vertebrate organism and propagated under conditions of physical containment comparable to PI and appropriate to the organism under study. It is important that the investigator demonstrate that the fraction of the viral genome being utilized does not lead to productive infection. A USDA permit is required for work with Class 5 agents.

III-B-4-b. For all experiments involving whole animals or plants and not covered by III-B-4-a, the appropriate containment will be determined by the IBC.

III-B-5. Experiments involving more than 10 liters of culture. The appropriate containment will be decided by the IBC. Where appropriate, the large-scale containment recommendations of the NIH should be used (45 FR24968).

III-C. Experiments that require institutional biosafety committee (IBC) notice simultaneously with initiation of experiments. Experiments not included in III-A, III-B, III-D and their subsections are to be considered in III-C. For example, experiments in which all components derive from nonpathogenic prokaryotes and nonpathogenic lower eukaryotes fall under III-C. All such experiments can be carried out at P1 containment. For experiments in this category, a registration document as described in III-B must be dated and signed by the investigator and filed with the local IBC. The IBC shall review all such proposals, but IBC review prior to initiation of the experiment is not required.

Caution: Experiments involving formation of recombinant DNA molecules containing no more than two thirds of the genome of any eukaryotic virus. Recombinant DNA molecules containing no more than two thirds of the genome of any eukaryotic virus (all viruses from a single family being considered identical) may be propagated and maintained in cells in tissue culture using P1 containment. For such experiments, it must be shown that the cells lack helper virus for the specific families of defective viruses being used. If helper virus is present, procedures specified under III-B.3 should be used. The DNA may contain fragments of the genome of viruses from more than one family but each fragment must be less than two thirds of a genome. (It is the responsibility of the institution and those associated with it to adhere to the intent of this Subpart and the NIH guidelines as well as to their specifics.)

III-D. Experiments exempt from the NIH guidelines and specified in section 61-1.3 of this Subpart. As provided in section 61-1.3(c) of this Subpart, an institution may be issued a certificate to engage in recombinant DNA activity limited to recombinant DNA molecules specified in section 61-1.3(c) (class III-D molecules) without being required to comply with the provisions of sections 61-1.30 and 61-1.31 of this Subpart pertaining to the establishment and functioning of an institutional biosafety committee (IBC).

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