Federal Regi^er / Vol. 47, No. 102 / Wednesday, May 26, 1982 / Notices 
23117 
that provide the means for replication of 
the recombinant DNAs and/or the host 
cells in which they replicate can be 
genetically designed to decrease by 
many orders of magnitude the 
probability of dissemination of 
recombinant DNAs outside the 
laboratory. Further details on biological 
containment may be found in Appendix 
1. 
As these three means of containment 
are complementary, different levels of 
containment appropriate for 
experiments with different recombinants 
can be established by applying various 
combinations of the physical and 
biological barriers along with a constant 
use of the standard practices. We 
consider these categories of 
containment separately in order that 
such combinations can be conveniently 
expressed in the Guidelines. 
In constructing these Guidelines, it 
was necessary to define boundary 
conditions for the different levels of 
physical and biological containment and 
for the classes of experiments to which 
they apply. We recognize that these 
definitions do not take into account all 
existing and anticipated information on 
special procedures that will allow 
particular experiments to be carried out 
under different conditions than 
indicated h^ without affecting risk. 
Indeed, we urge that individual 
investigators devise simple and more 
effective containment procedures and 
that investigators and institutional 
biosafety committees recommend 
changes in the Guidelines to permit their 
use. 
III. Containment Guidelines for Covered 
Experiments 
Part III discusses experiments 
involving recombinant DNA. These 
experiments have been divided into four 
classes; 
UI-A. Experiments which require 
specific RAC review and NIH and 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 procedures of the Guidelines. 
If an experiment falls into both Class 
III-A and one of the other clases, 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 
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, NIH. (See Sections IV- 
C-l-b-(l), IV-C-l-b-(2), and 
subsections.) 
III-A. Experiments that Require RAC 
Review and NIH and IBC 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 Registn 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-J. Deliberate formation of 
recombinant DNAs containing genes for 
the biosynthesis of toxic molecules 
lethal for vertebrates at an LDso of less 
than 100 nanograms per kilogram body 
weight (e.g., microbial toxins such as the 
botidinum 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 Appdenix F. 
IU-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 (2), 
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 IBC 
Approval Before Initiation. Investigators 
performing experiments in this category 
must submit to their Institutional 
Biosafety Committee (IBC), prior to 
initiation of the exq)eriments, 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 
these Guidelines. This registration 
document must be dated and signed by 
the investigator and filed only with the 
local IBC. The IBC shall review ail 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-(3j.) 
in-B-1. Experiments Using Human or 
AnimpI Pathogens (Class 2, Class 3, 
Class 4, or Class 5 Agents [1]) as Host- 
Vector Systems. 
ni-B-l-a. Experiments involving the 
introduction of recombinant DNA into 
Class 2 agents can be carried out at P2 
containment. 
III-B-l-b. Experiments involving the 
introduction of recombinant DNA into 
Class 3 agents can be carried out at P3 
containment. 
III-B-l-c. Experiments involving the 
introduction of recombinant DNA into 
Class 4 agents can be carried out at P4 
contain m ent. 
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 USDA 
permit is required for work with Class 5 
agents [18, 20). 
in-B-2. Experiments in Which DNA 
from Human or Animal Pathogens 
(Class 2, Class 3^ Class 4, or Gass S' 
Agents [If) 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 tranferred into 
nonpathogenic puokaryotes or lower 
eukaryotes may be performed under P2 
containment. Recombinant DNA 
experiments in which DNA fix>m €3ass 4 
agents is transferred into nonpathogenic 
prokaryotes or lower eukaryotes can be 
pm'formed at P2 containment after 
demonstration that only a totally and 
irreversibly defective fraction of the 
agenf s viral genome is present in a 
given recombinant. In the absence of 
such a demonstration, P4 containment 
should be used. Specific lowering of 
containment to Pi for particular 
experiments can be approved by the 
IBC. Many experiments in this categcHry 
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 requires RAC review and 
NIH approval (see Section III-a-1), or 
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