NOTICES 
33119 
more (or less) safe than seen at this 
time and that the currently assigned 
containment level should be changed. 
Therefore the inserted passage states 
that “changes in these levels for spe- 
cific experiments (or the assignment 
of levels to experiments not explicitly 
considered in this section) may be ex- 
pressly approved by the Director, NITI, 
on the recommendation of the Recom- 
binant DNA Advisory Committee.” 
Permissible experiments using E. Coli 
K-l 2 host-vector systems 
Eukaryotic DNA Sources. There was 
disagreement over those provisions in 
the Guidelines that allow the princi- 
pal investigator to choose between two 
combinations of containment proce- 
dures. In several instances, for exam- 
ple, one is permitted to use P2 + EK1 
or P1 + EK2. 
This flexibility provision was en- 
dorsed by some commentators but 
questioned by others. It was discussed 
above in Part II. This concept of inves- 
tigator flexibility is not a new one; it 
was allowed under the original Guide- 
lines. Based upon events of the past 
two years, the RAC merely proposed 
that the principle be extended to cer- 
tain specified additional cases where 
they believe it appropriate. Included 
in the PRG-NIH are all such specific 
cases of flexibility recommended in 
the PRG-RAC. 
On the other hand, in certain other 
specific cases (e.g., DNA from birds), 
the PRG-RAC recommended that the 
containment level be P2+EK2, with- 
out the option of P3+EK1. Certain 
commentators urged that in all cases 
where the containment level of 
P2+EK2 is given, the option of 
P3 + EK1 be allowed. However, the 
RAC felt that in view of their in- 
creased confidence in the biological 
containment offered by the EK2 
system, P2 + EK2 offers more contain- 
ment than P3+EK1, and that 
P2+EK2 without the option of 
P3+EK1 should be the containment 
level for certain specified classes of ex- 
periments. Therefore, there is speci- 
fied in the PRG-NIH the containment 
levels of P2+EK2 without the option 
of P3+EK1 in every case where it ap- 
peared in the PRG-RAC. 
Discussed below and in the accompa- 
nying “Decision” document is the reas- 
sessment which was made of the clon- 
ing of viral DNA into E. coli K-12 at 
the Ascot Workshop and the April 6-7, 
1978, Working Group meeting that en- 
dorsed the Ascot report. The RAC at 
its April 27-28 meeting unanimously 
endorsed the Working Group report 
recommending lower containment 
levels for deliberate cloning of viral 
DNA into E. coli K-12. One of the rea- 
sons given originally for the higher 
containment level for shotgun experi- 
ments involving primate DNA into E. 
coli K-12 was the possible inadvertant 
cloning of viral DNA. In view of their 
recommendation of lower containment 
for deliberate cloning of viral DNA 
into E. coli K-12, the RAC on April 
27-28, 1978, reconsidered primate shot- 
gun levels and voted unanimously for 
new language as follows; "Primates. 
P2 physical containment + an EK2 
host-vector. Any lowering of contain- 
ment below these levels (i.e., for puri- 
fied DNA or characterized clones) 
cannot be made solely by an institu- 
tional biosafety committee but re- 
quires NIH approval.” This new lan- 
guage is inserted in the PRG-NIH, as 
well as a similar lowering of contain- 
ment for shotgun cloning of cold- 
blooded vertebrate DNA into E. coli 
K-12. 
Prokaryotic DNA Sources. In the 
1976 guidelines the section dealing 
with shotgun experiments in which 
prokaryotic DNA is inserted into E. 
coli K-12 was subdivided into two 
parts— “Prokaryotes That Exchange 
Genetic Information with E. coli” and 
“Prokaryotes That Do Not Exchange 
Genetic Information with E. coli. ” In 
the PRG-RAC it was assumed that all 
prokaryotes that exchange genetic in- 
formation with E. coli would be 
exempt from the guidelines by appear- 
ing on the “list of non-novel exchang- 
ers.” Therefore, the PRG-RAC the 
section dealing with these experiments 
actually considered only prokaryotes 
that did not exchange genetic infor- 
mation with E. coli. The problem with 
this approach was discussed by com- 
mentators, focusing especially on the 
case of Agrobacterium tumefaciens. It 
meant that a prokaryote which ex- 
changes genetic information with E. 
coli, and was therefore properly as- 
signed a low containment level under 
the 1976 guidelines, would under the 
PRG-RAC either appear on the list 
and be exempt from the guidelines or, 
if not appearing on the list for some 
reason, would require in some cases a 
higher containment. This was not the 
intent of the RAC. Therefore, the 
RAC agreed at their April 27-28 meet- 
ing that language should be reinserted 
into the PRG-NIH covering prokar- 
yotes that exchange genetic informa- 
tion with E. coli but do not appear on 
the list, and this has now been done. 
For prokaryotes that do not ex- 
change genetic information with E. 
coli the PRG-RAC proposed that 
P1 + EK2 or P2+EK1 conditions apply 
only in cases of extensive characteriza- 
tion and RAC approval. A number of 
commentators objected. Some felt 
that experiments involving non- 
pathogenic prokaryotes should be con- 
ducted at these lower levels without 
extensive characterization or RAC ap- 
proval, and others argued that plant 
pathogens should not be included with 
CDC class 2 agents as requiring 
P3+EK2 containment. The RAC at 
their April meeting agreed with the 
commentators. Accordingly, this sec- 
tion of the PRG-NIH has been rewrit- 
ten. 
The EMBO Standing Advisory Com- 
mittee on Recombinant DNA Re- 
search recommends that the contain- 
ment level for all experiments involv- 
ing the insertion of novel nonpatho- 
genic prokaryotic DNA into E. coli K- 
12 be P1 + EK1. Acting conservatively, 
the Director has retained in the PRG- 
NIH the levels of P2 + EK1 or EK2 for 
nonpathogenic prokaryotes that do 
not exchange genetic information with 
E. coli. 
The PRG-RAC received substantial 
criticisms for identifying all agents 
classified as class 2 in the CDC’s publi- 
cation “Classification of Etiologic 
Agents on the Basis of Hazard” 
(fourth edition, July 1974) as being 
pathogenic for the purpose of assign- 
ing containment levels. Several com- 
mentators stated that many of the or- 
ganisms so classified were harmless 
and that others were of such low path- 
ogenicity that severe safety precau- 
tions were unwarranted. It was also 
pointed out that the pathogenicity of 
an intact microorganism and the con- 
jectural hazard of a piece of DNA 
from such an organism with E. coli K- 
12 were quite different matters. The 
suggestion of these commentators has 
been accepted, and thus footnote 1 has 
been added to the PRG-NIH. This 
gives NIH the authority, upon the rec- 
ommendation of the RAC, to consider 
certain class 2 agents as class 1 for the 
purpose of these guidelines. 
Plasmid, Phage, and Virus DNA 
Sources. Many of the commentators 
agreed that both the original guide- 
lines and the PRG-RAC were overly 
stringent with regard to virus experi- 
ments. In commenting on the PRG- 
RAC, the EMBO Standing Advisory 
Committee on Recombinant DNA Re- 
search wrote, “The EMBO Committee 
believes that the containment categor- 
ization of experiments with animal 
virus DNA’s which is proposed by the 
NIH Advisory Committee is too indis- 
criminate and excessively stringent 
considering the proposed classification 
of experiments with other classes of 
DNA and the longstanding, accepted 
safety precautions for handling intact 
virus particles and viral nucleic acids 
* * *.” The EMBO Committee pro- 
posed (1) that experiments with viral 
DNA be considered on a case-by-case 
basis or (2) that a detailed set of rec- 
ommended categories for such experi- 
ments be produced. 
A joint United States-EMBO Work- 
ship To Assess Risks for Recombinant 
DNA Experiments Involving the Gen- 
omes of Animal, Plant, and Insert Vir- 
uses was held in Ascot, England, or. 
January 26-28, 1978. The workship 
FEDERAL REGISTER, VOL. 43, NO. 146— FRIDAY, JULY 28, 1978 
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