462G8 
Federal Register / Vol. 49, No. 227 / Friday, November 23, 1984 / Notices 
Before Initiation. Experiments in this 
category cannot be initiated without 
submission of relevant information on 
the proposed experiment to N1H, 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). 
III-A-1. Deliberate formation of 
recombinant DNAs containing genes for 
the biosynthesis of toxic molecules 
lethal for vertebrates at an LD«, of less 
than 100 nanograms per kilogram body 
weight (e.g., microbial toxins such as the 
botulinum toxins, tetanus toxin, 
diptheria toxin, Shigella dysenteriae 
neurotoxin). Specific approval has been 
given for the cloning in E. coli K-12 of 
DNA 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. 
Ill-A-2. Deliberate release into the 
environment of any organism containing 
recombinant DNA, except certain plants 
as described in Appendix L 
III-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 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 
submission 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) th° 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 Guidlelines. 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)). 
IIl-B-1. Experiments Using Human or 
Animal Pathogens (Class 2, Class 3, 
Class 4, or Class 5 Agents (1) as Host- 
Vector Systems. 
III-B-l-a. Experiments involving the 
introduction of recombinant DNA into 
Class 2 agents can be carried out at BL2 
containment. 
HI-B-l-b. Experiments involving the 
introduction of recombinant DNA into 
Class 3 agents can be carried out at BL3 
containment. 
III-B-l-c: Experiments involving the 
introduction of recombinant DNA into 
Class 4 agents can be carried out at BL4 
containment. 
III-B-l-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 U.S. 
Department of Agriculture (USDA) 
permit is required for work with Class 5 
agents [18, 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 (7) 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 to 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. 
UI-B-2-h. Containment conditions for 
experiments in which DNA from Class 5 
agents is transferred into nonpthogenic 
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 
[18, 20). 
IIl-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(77) being considered identical (75) 
may be considered defective and can be used 
in the absence of helper under the conditions 
specified in Section III— C. 
III-B-3-a. Experiments involving the 
use of infectious Class 2 animal viruses 
(7) or defective Class 2 animal viruses in 
the presence of helper virus can be 
performed at BL2 containment. 
IlI-B-3-b. Experiments involving the 
use of infectious Class 3 animal viruses 
(7) or defective Class 3 animal viruses in 
the presence of helper virus can be 
carried out at BL3 containment. 
III-B-3-c. Experiments involving the 
use of infectious Class 4 viruses (7) or 
defective Class 4 viruses in the presence 
of helper virus may be carried out under 
BL4 containment. 
III-B-3-d. Experiments involving the 
use of infectious Class 5 (7) 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 [18, 20). 
lll-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 Sections III-B-3-a, III— B — 3— b. III— B— 
3-c, or III— B— 3 — d may be carried out 
under BLl containment. 
I1I-B-4. Recombinant DN. 
Experiments Involving Whole Animals 
or Plants. 
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