42914 
Federal Register / Vol. 44, No. 141 / Friday, July 20, 1979 / Notices 
DEPARTMENT OF HEALTH, 
EDUCATION, AND WELFARE 
National Institutes of Health 
Recombinant DNA Research; Actions 
Under Guidelines 
AGENCY: National Institutes of Health. 
action: Notice of actions under NIH 
Guidelines for Research Involving 
Recombinant DNA Molecules. 
SUMMARY: This notice sets forth actions 
taken by the Director, NIH under the 
1978 NIH Guidelines for Research 
Involving Recombinant DNA Molecules 
(43 FR 60108). 
EFFECTIVE DATE: July 20, 1979. 
FOR FURTHER INFORMATION CONTACT: 
Additional information can be obtained 
from the Office of Recombinant DNA 
Activities, National Institutes of Health, 
Bethesda, Maryland 20205, (301) 496- 
6051. 
SUPPLEMENTARY INFORMATION: I am 
promulgating today several major 
actions under the NIH Guidelines for 
Research Involving Recombinant DNA 
Molecules. These proposed actions were 
published for comment in the Federal 
-Register of April 13, 1979 and reviewed 
and recommended for approval by the 
Recombinant DNA Advisory Committee 
(RAC) at its meeting on May 21, 22, and 
23, 1979. In accordance with Section IV- 
E-l-b of the NIH Guidelines, I find that 
these actions comply with the 
Guidelines and present no significant 
risk to health or the environment. 
Part I of this announcement provides 
background information on the actions. 
Part II provides a summary of the major 
actions. 
I. Decisions on Actions Under 
Guidelines 
A. Containment levels for certified 
Saccharomyces Cerevisiae and 
Neurospora Crassa HV systems 
The RAC at its February 15-16, 1979 
meeting recommended the use of 
Saccharomyces cerevisiae and 
Neurospora crassa as HVl systems and 
specified certain strains and vectors of 
S. cerevisiae as HV2 host-vector 
systems. The certified systems were to 
be used as follows: 
‘‘In accord with Section III — C— 5, host-vector 
systems which have been approved as HVl 
systems may be used under P2 containment 
conditions for shotgun experiments with 
phages, plasmids, and DNA from 
nonpathogenic prokaryotes which do not 
produce polypeptide toxins (34). For other 
classes of recombinant DNA experiments 
with these HVl systems, except for the 
cloning of complete genomes of eukaryote 
viruses, the S. cerevisiae and N. crassa HVl 
systems and S. cerevisiae HV2 systems may 
be used at the physical containment levels 
applicable to EK1 and EK2 systems, 
respectively." 
While the RAC generally approved 
equivalence between these HV systems 
and the EK designations in the 
Guidelines, there was concern raised 
about the appropriate levels of 
containment when complete genomes of 
eukaryotic viruses are cloned into these 
organisms. 
Prior to the RAC considering this 
issue at it February 15-16, 1979 meeting, 
it was mistakenly printed in the Federal 
Register of January 15, 1979 as follows: 
“Experiments involving complete genomes 
of class 1 eukaryote viruses will require P3 + 
HV2 containment levels. Other eukaryote 
viruses are to be handled on a case-by-case 
basis [45].’’ 
The correct wording should have been 
‘‘will require P3 + HVl or P2 + HV2 
containment levels.” 
The RAC in its deliberations at the 
February 15-16, 1979 meeting took a less 
restricted position and recommended 
that the following wording be 
substituted for the previous proposal: 
“Experiments involving complete genomes 
of eukaryote viruses will require P3 + HVl 
or P2 + HV2 containment levels." 
This recommendation passed by a 
vote of 17 to 1 with 2 abstentions. Since 
this was a major change in the proposed 
action as it appeared in the Federal 
Register on January 15, 1979, additional 
opportunity for public comment was 
deemed appropriate. Accordingly, this 
proposal was published in the Federal 
Register on April 13, 1979 for an 
additional 30-day comment period prior 
to its presentation at the May 21-23, 
1979 RAC meeting. During the 30-day 
comment period no comments were 
received. 
Based on the containment features of 
these organisms which were previously 
presented in reports to the RAC and the 
discussion at the February 15-16, 1979 
meeting, the RAC at its May, 1979 
meeting voted 17 to 0 with 2 abstentions 
to accept this proposal. 
B. Containment levels for unmodified 
laboratory strains of Neurospora Crassa 
Based on extensive analysis of the 
fungus N. crassa, the NIH has previously 
approved genetically modified strains of 
this organism as HVl as stated in the 
Federal Register of April 11, 1979. Atjts 
February 15-16, 1979 meeting, the RAC 
also recommended a limited use of 
unmodified laboratory strains of N. 
crassa. The NIH accepted the following 
conservative interpretation of the RAC’s 
position based on the discussion at the 
time and until there was opportunity for 
further clarification by the RAC: 
"Unmodified laboratory strains of N. 
crassa are approved at the P3 level of 
containment for shotgun experiments with 
phages, plasmids, and DNA from Class 1 
prokaryotes (1) and lower eukaryotes that do 
not produce polypeptide toxins |34].“ 
Based on the need for further 
clarification, the following alternate 
interpretation of the RAC’s action was 
published in the Federal Register on 
April 13, 1979 for an additional 30-day 
comment period prior to its 
consideration at the May 21-23, 1979 
meeting. During the 30-day comment 
period no comments were received on 
this proposal: 
"Unmodified laboratory strains of 
Neurospora crassa can be used in all 
experiments for which HVl N. crassa 
systems are approved provided that these are 
carried out at physical containment one level 
higher than required for HVl. However, if P3 
containment is specified for HVl N. crassa, 
this level is considered adequate for 
unmodified N. crassa. For P2 physical 
containment, special care must be exercised 
to prevent aerial dispersal of macroconidia, 
including the use of a biological safety 
cabinet." 
The discussion on this issue at the 
May, 1979 RAC meeting followed that 
which has been described in the Federal 
Register of April 11, 1979. Essentially, 
there was cfencern about the escape of 
N. crassa since it forms spores which 
are freely dispersed. As a result of this 
concern, it was recommended at the 
February, 1979 meeting that all 
experiments with wild type N. crassa 
should require the use of a biological 
safety cabinet. At the May 21-23, 1979 
meeting of the RAC, it was pointed out 
that the organism has ony a small 
ecological niche and that it is a 
nonpathogenic organism. 
The RAC accepted the use of 
unmodified laboratory strains of N. 
crassa as published in the Federal 
Register of April 13, 1979, by a vote of 11 
to 2 with 5 abstentions. It was the sense 
of the RAC that the principle of 
equivalency of HV systems with EK 
systems applies at the present time only 
to the setting of containment levels for 
shotgun experiments. It does not apply 
at the present time to lowering of 
containment levels for characterized or 
purified DNA preparations and clones, 
to returing DNA segments to non-HVl 
host of origin, etc. 
C.l Transfer of cloned DNA segments to 
Eukaryotic organisms 
Based on the recommendation of the 
RAC at its February 15-16, 1979 meeting, 
the NIH previously has approved the 
[ 128 ] 
