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Federal Register / Vol. 51. No. 88 / Wednesday. May 7, 1986 / Notices 
exempt from the requirements of the 
Guidelines. 
Changes in containment levels from 
those specified here may not be 
instituted without the express approval 
of the Director, N1H (see Sections IV-C- 
l-b-(l), IV — C— 1— b— (2). and subsections). 
Ill- A — Experiments That Require RAC 
Review and NIH and IDC Approval 
Before Initiation 
Experiments in this category cannot 
be initiated without submission of 
relevant information on the proposed 
experiment to NIH. the publication of 
the proposal in the Federal Register for 
thirty days of comment, review by the 
RAC, and specific approval by NIH. The 
containment conditions for such 
experiments will be recommended by 
RAC and 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 section and the appropriate 
containment conditions are listed in 
Appendices D and F. If an experiment is 
similar to those listed in Appendices D 
and F. ORDA may determine 
appropriate containment conditions 
according to case precedents under 
Section IV-C-l-b-(3)-(g). 
If the experiments in this category are 
suomitted for review to another Federal 
agency, the submitter shall notify 
CEDA; ORDA may then determine that 
such review serves the same purpose, 
and based on that determination, notify 
the submitter that no RAC review will 
take place, no NIH approval is 
necessary, and the experiment may 
proceed upon approval from the other 
Federal agency. 
I1I-A-1. Deliberate formation of 
recombinant DNAs containing genes for 
the biosynthesis of toxic molecules 
lethal for vertebrates at an LD 5 o 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 ICO 
nar.ograms to 100 micrograms per 
kilogram body weight. Containment 
levels for these experiments are 
specified in Appendix F. 
III-A-2. Deliberate release into the 
environment of any organism containing 
recombinant DNA, except certain plants 
as described in Appendix L. 
I1I-A-3. Deliberate transfer of a drug 
resistance trait to microorganisms that 
are not known to acquire it naturally [2], 
if such acquisition could compromise the 
use of the drug to control disease agents 
in human or veterinary medicine or 
agriculture. 
III-A-4. Deliberate transfer of 
recombinant DNA or DNA or RNA 
derived from recombinant DNA into 
human subjects [21]. The requirement 
for RAC review should not be 
considered to preempt any other 
required review of experiments with 
human subjects. Institutional Review 
Board (IRB) review of the proposal 
should be completed before submissin to 
NIH. 
III-B — Experiments That Require IBC 
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: (i) The source(s) of DNA: 
(ii) the nature of the inserted DNA 
sequences: (iii) the hosts and vectors to 
be used: (iv) whether a deliberate 
attempt will be made to obtain 
expression of a foreign gene, and, if so, 
what protein will be produced; and (v) 
the containment conditions specified in 
these 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 (see 
Section IV-C-l-b-(3)). 
III-B-1 — Experiments Using Human or 
Animal Pathogens (Class 2, Class 3, 
Class 4, or Class 5 A.gents [ 1 ]) as Host- 
Vector Systems 
lll-B-l-a. Experiments involving the 
introduction of recombinant DNA into 
Class 2 agents can be carried out at BL2 
containment. 
III-B-l-b. Experiments invoking the 
introduction of recombinant DNA into 
Class 3 agent3 can be carried out at BL3 
containment. 
lll-B-l-c. Experiments involving the 
introduction of recombinant DNA into 
Class 4 agents can be carried out at BL4 
containment. 
111-B-l-d. Containment conditions for 
experiments involving the introduction 
of recombinant DNA into Class 5 agents 
wili be set on a case-by-case basis 
following ORDA review. A U.S. 
Department of Agriculture (USDA) 
permit is required for work with Class 5 
agents ri8, 20]. 
III-B-2 — Experiments in Which DNA 
From Human or Animal Pathogens 
(Class 2. Class 3, Class 4, or Class 5 
Agents [1]) 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 [1] is transferred into 
nonpathogenic prokaryotes or lower 
eukaryotes may be performed under BL2 
containment. Recombinant DNA 
experiments in which DNA from Class 4 
agents is transferred into nonpathogenic 
prokaryotes or lower eukaryotes can be 
performed at BL2 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. BL4 containment should 
be used. Specific lowering of 
containment of BLl for particular 
experiments can be approved by the 
IBC. Many experiments in this category 
will be exempt from the Guidelines (see 
Sections III— D — 4 and III— D— 5). 
Experiments involving the formation of 
recombinant DNAs for certain genes 
coding for molecules toxic for 
vertebrates require RAC review and 
NIH approval (see Section III— A— 1) or 
must be carried out under NIH specified 
conditions as described in Appendix F. 
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 
[13. 20 J. 
III-B-3 — Experiments Involving the Use 
of Infectious Animal or Plant DNA or 
RNA Viruses or Defective Animal or 
Plant DNA or RNA 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 or 
RNA molecules derived therefrom, which 
contain less than two-thirds of the genome of 
any eukaryotic virus (all virus from a single 
Family (17] being considered identical [19]). 
may be considered defective and can be used 
[ 4 ] 
Recombinant DNA Research, Volume 1 1 
