Federal Register / Vol. 59, No. 127 / Tuesday, July 5, 1994 / Notices 
34477 
experiments in this category prior to 
their initiation. Requests to decrease the 
level of containment specified for 
experiments in this category will be 
considered by NIH (see Section IV-C-i- 
M2Hc)). 
Section IiI-C-1. Experiments Using 
Human or Animal Pathogens (Class 2, 
Class 3, Class 4, or Class 5 Agents (See 
Section V-A) as Host-Vector Systems 
Section lll-C-l-a. Experiments 
involving the introduction of 
recombinant DNA into Class 2 agents 
shall be conducted at Biosafety Level 
(BL) 2 containment. Experiments with 
such agents shall be conducted with 
whole animals at BL2 or BL2-N 
(Animals) containment. 
Section Ill-C-l-b. Experiments 
involving the introduction of 
recombinant DNA into Class 3 agents 
shall be conducted at BL3 containment. 
Experiments with such agents shall be 
conducted with whole animals at BL3 or 
BL3-N containment. 
Section Ill-C-l-c. Experiments 
involving the introduction of 
recombinant DNA into Class 4 agents 
shall be conducted at BL4 containment. 
Experiments with such agents shall be 
conducted with whole animals at BL4 or 
BL4-N containment. 
Section 111-C-l-d Containment 
conditions for experiments involving 
the introduction of recombinant DNA 
into Class 5 agents shall be set on a case- 
by-case basis following NIH/ORDA 
review. A U S. Department of 
Agriculture permit is required for work 
with Class 5 agents (see Sections V-R 
and V-T). Experiments with such agents 
shall be conducted with whole animals 
at BL4 or BL4-N containment 
Section Ill-C-2. Experiments in Which 
DNA From Human or Animal Pathogens 
(Class 2, Class 3, Class 4, or Class 5 
Agents (See Section V-A) is Cloned Into 
Nonpathogenic Prokaryotic or Lower 
Eukaryotic Host-Vector Systems 
Section llI-C-2-a. Experiments in 
which DNA from Class 2 or Class 3 
agents (see Section V-A) is transferred 
into nonpathogenic prokaryotes or 
lower eukaryotes may be performed 
under BL2 containment. Experiments in 
which D.NA from Class 4 agents is 
transferred into nonpathogenic 
prokaryotes or lower eukaryotes may be 
performed under 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 shall 
be used. The Institutional fLosafety 
Committee may approve ifie spe< ifi«. 
lowering of f.ontainmeni 'or partK.ular 
experiments to BLl. Many experiments 
in this category are exempt from the 
NIH Guidelines (see Section IIl-E). 
Experiments involving the formation of 
recombinant DNA for certain genes 
coding for molecules toxic for 
vertebrates require NIH/ORDA approval 
(see Section IlI-B-1) or shall be 
conducted under NIH specified 
conditions as described in Appendix F. 
Section III-C-2-b. Containment 
conditions for experiments in which 
DNA horn Class 5 agents is transferred 
into nonpathogenic prokaryotes or 
lower eukaryotes shall be determined by 
NIH/ORDA following a case-by-case 
review. A U.S. Department of 
Agriculture permit is required for work 
with Class 5 agents (see Sections V-R 
and V-T). 
Section lII-C-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 that permit a 
productive infection. In such cases, 
serious consideration should be given to 
increasing physical containment by at 
least one level. 
Note: Recombinant DNA or RNA molecules 
derived therefrom, which contain less than 
two-thirds of the genome of any eukaryotic 
virus (all vinises from a single Family (see 
Section V-Q) being considered identical (see 
Section V-S), are considered defective and 
may be used in the absence of helper under 
the conditions specified in Section III-D-1. 
Section llI-C-3-a. Experiments 
involving the use of infectious or 
defective Class 2 animal viruses (see 
Section V-A, Appendix B-11, and 
Appendix B-ll-E) in the presence of 
h^per virus may be conducted at BL2. 
Section IIl-C-3-b. Experiments 
involving the use of infectious or 
defeiTtive Class 3 animal viruses (see 
Section V-A and Appendix B-llI-D) in 
the presence of helper virus may be 
conducted at BL3. 
Section III-C-3-c. Experiments 
involving the use of infectious or 
defective Class 4 animal viruses (s£*e 
Section V-A and Appendix B-IV-D) in 
the presence of helper virus may be 
conduded at BL4. 
Section Ill-C-3-d. Experiments 
involving the use of infectious or 
defective Class 5 viruses (see Section V- 
A and Appendix B V) in the presence 
of helper virus shall be determined on 
a case-by-case basis following NIH/ 
ORDA review. A U.S. Department of 
Agriculture permit is required for work 
with Class 5 agents (see Sections V-R 
and V-T). 
Section III-C-3-e. Experimeats 
involving the use of infectious or 
defective animal or plant viruses in the 
presence of helper virus are not covered 
in Sections III-C-3-a through lll-C-3- 
d and may be conducted at BLl. 
Section lII-C-4. Experiments Involving 
Whole Animals 
This section covers experiments 
involving whole animals in which the 
animal’s genome has been altered by 
stable introduction of recombinant 
DNA, or DNA derived therefrom, into 
the germ-line (transgenic animals) and 
experiments involving viable 
recombinant DNA-modified 
microorganisms tested on whole 
animals. For the latter, other than 
viruses which are only vertically 
transmitted, the experiments may not be 
conducted at BLl-N containment. A 
minimum containment of BL2 or BL2- 
N is required. 
Caution — Special care should be used 
in the evaluation of containment 
conditions for some experiments with 
transgenic animals. For example, such 
experiments might lead to the creation 
of novel mechanisms or increased 
transmission of a recombinant pathogen 
or production of undesirable traits in 
the host animal. In such cases, serious 
consideration should be given to 
increasing the containment conditions. 
Section IIl-C— 4-a. Recombinant DNA. 
or DNA or RNA molecules derived 
therefrom, from any source except for 
greater than two-thirds of eukaryotic 
viral genome may be transferred to any 
non-human vertebrate or any 
invertebrate organism and propagated 
under conditions of physical 
containment comparable to BLl o: BLl- 
N and appropriate to the organism 
under study (see Section V-B). 
Animals that contain sequences from 
viral vectors, which do not lead to 
transmissible infection either directly or 
indirectly as a result of 
complementation or recombination in 
animals, may be propagated under 
conditions of physical containment 
comparable to BLl or BLl-N and 
appropriate to the organi.sm under 
study. Experiments involving the 
introduction of other sequences from 
eukaryotic viral genomes into animal.^ 
are covered under Section lIl-C-4-b. 
For experiments involving recomhiiianl 
DNA-modified Class 2, 3, 4, or 5 
organisms, stje Section V-A. It is 
iiri|)ortant that the investigator 
Recombinant DNA Research, Volume 19 
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