Federal Register / Vol. 44, No. 141 / Friday, July 20, 1979 / Notices 
42915 
return of DNA segments to a higher 
eukaryotic host of origin as stated in the 
Federal Register on April 11, 1979: 
"IU-C-6. Return of DNA Segments to a 
Higher Eukaryotic Host of Origin. DNA from 
a higher eukaryote (Host D) may be inserted 
into a lambdoid phage vector or into a vector 
from certified EK2 host-vector system and 
propagated in E. coli K-12 under the 
appropriate containment conditions [see 
Section III— A— 1). Subsequently, this 
recombinant DNA may be returned to Host D 
and propagated under conditions of physical 
containment comparable to Pi and 
appropriate to the organism under study 
|2A|." 
Several commentators had requested 
that this section be broadened to permit 
the heterologous transfer of DNA 
segments to a eukaryote other than the 
host of origin. A broader proposal could 
not be considered at the February 15-10, 
1979 RAC meeting as it would require 
the opportunity for public comment. The 
following proposed revision of Section 
IU-C-6 was published in the Federal 
Register on April 13, 1979 for a 30-day 
comment period prior to its 
consideration at the May 21-23, 1979: 
"III-C-6. Transfer of Cloned DNA 
Segments to Eukaryotic Organisms. DNA 
from any nonprohibited source [Section 1-D) 
which has been cloned and propagated in E 
coli under appropriate physical containment 
conditions, may be transferred with the E 
coli vector used for cloning to a ekaryotic 
organism or cells in culture and propagated 
under conditions of physical containment 
comparable to Pi and appropriate to the 
organism under study [2A]." 
Several letters were received during 
the comment period which supported 
this proposal. Recent experimental 
results have demonstrated that it is 
possible to transfer DNA to eukaryotic 
organisms without a requirement for the 
DNA to be part of a recombinant DNA 
molecule. The major difference in the 
interspecies experiments which involve 
using recombinant DNA techniques is 
that there will be an association of the 
E. coli vector DNA with the DNA of 
interest. The recombinant DNA methods 
also allow a more controlled process 
since it permits the use of selected 
genes. 
A very important category of 
experiments which this amendment 
would permit involves the cloning of 
DNA from one higher eukaryote into E. 
coli. followed by the transfer to an 
embryo or teratoma of another 
eukaryote. This procedure wiU enable 
the study of the genetic basis of various 
diseases by isolating individual genes 
and examining their expression in 
various whole animals. There will be the 
possibility of understanding the basis 
for cell diversification during 
development of higher organisms and 
the organization of genetic information. 
These features may be important in 
many cases to understanding the origins 
of malignant growth and the genetic 
basis of disease. As noted by one 
commentator, this research may lead to 
the “possible cure of human genetic 
diseases.” 
One commentator indicated that this 
proposal would, in essence, allow for 
nearly any eukaryote to become a host 
for any DNA: this would not be in the 
spirit of the Guidelines. Another 
commentator noted that any 
experiments that involved the return of 
cloned DNA to humans would require 
the examination by human 
experimentation committees. 
These comments were discussed at 
the May 21-23. 1979 RAC meeting, and 
concern was expressed over the broad 
nature of the proposal. The use of 
recombinant DNA methods for studying 
diseases using whole animals or plants 
was generally supported by the RAC. It 
was agreed that this revision should 
appear as a new section of the 
Guidelines, III-C-7. The following more 
restrictive amendment was proposed by 
the RAC to limit the experiments to 
easily contained whole organisms and 
to only small portions of viruses: 
"LU-C-7. Transfer of Cloned DNA 
Segments to Eukaryotic Organisms. 
IIl-C-7-a. Transfer to Non-human 
Vertebrates. DNA from any nonprohibited 
source [Section I-D], except for greater than 
one quarter of a eukaryotic viral genome, 
which has been cloned and propagated in E. 
coli under appropriate physical containment 
conditions, may be transferred with the E. 
coli vector used for cloning to any eukaryotic 
cells in culture or to any non-human 
vertebrate organism and propagated under 
conditions of physical containment 
comparable to Pi and appropriate to the 
organism under study [2AJ. Transfers to any 
other host will be considered by the RAC on 
a case-by-case basis [45]. 
IlI-C-7-b. Transfer to Higher Plants. DNA 
from any nonprohibited source [Section 1-D] 
which has been cloned and propagated in E 
coli under appropriate containment 
conditions, may be transferred with the E. 
coli vector used for cloning to any higher 
plant organisms (Angiosperms and 
Gymnosperms) and propagated under 
conditions of physical containment 
comparable to Pi and appropriate to the 
organism under study [2A]. Intact plants or 
propagative plant parts may be grown under 
Pi conditions described under Section III— C— 
3. Containment must be modified to ensure 
that the spread of pollen, seed or other 
propagules is prevented. This can be 
accomplished by conversion to negative 
pressure in the growth cabinet or greenhouse 
or by physical entrapment by "bagging” of 
reproductive structures. Transfers to any 
[ 129 ] 
other plant organisms will be considered on a 
case-by-case basis [45]." 
The RAC accepted proposal III-C-7-a 
by a vote of 19 to 2 with 2 abstentions, 
and proposal IU-C-7-b by a vote of 18 
to 0 with 1 abstention. 
D. Proposed exemption under I-E-5 for 
cloning in tissue culture cells 
The RAC considered a proposal for 
exempting experiments involving the 
propagation of recombinant DNA 
molecules from non-viral components in 
tissue culture cells. The proposal, made 
by Dr. Wallace Rowe, appeared in the 
April 13, 1977 Federal Register as 
follows: 
"Those recombinant DNA molecules that 
are propagated in cells in tissue culture and 
that are derived entirely from non-viral 
components (that is, no component is derived 
from a eukaryotic virus) or that contain no 
more than one-fourth of the genome of a 
eukaryotic virus are exempt from the 
Guidelines." 
During the 30-day period for comment, 
one comment was received on the 
proposed action. This commentator 
opposed the motion on the grounds that 
the introduction of recombinant DNA 
molecules linked even to only one-fourth 
of a viral genome in tissue culture cells 
may possibly generate altered 
endogenous or exogenous viruses in the 
cells. 
The RAC considered this proposal 
following a discussion of its merits by 
Dr. Rowe. It was pointed out that tissue 
culture cells are well contained and safe 
systems for studying gene function. 
Some members of the RAC expressed 
concern that recombinant molecules 
containing one-fourth of a viral genome 
might possibly generate infectious virus 
Dr. Rowe agreed to amend his proposal 
to delete the portion that referred to 
exempting recombinant molecules 
containing less than one-fourth of a 
eukaryotic viral genome. 
The RAC voted 17 to 3 with 2 
abstentions to accept the amended 
proposal with a minor modification in 
the wording to include "and 
maintained" in cells, as follows: 
"Those recombinant DNA molecules that 
are propagated and maintained in cells in 
tissue culture and that are derived entirely 
from non-viral components (that is, no 
component is derived from a eukaryotic 
virus) are exempt from the Guidelines." 
E. Containment levels for experiments 
involving Genera Streptomyces and 
Micromonospora 
The RAC at its May 21-23, 1979 
meeting considered a proposal that had 
been submitted by the Working Group 
on Prokaryotic Host-Vectors other than 
E. coli that would allow the formation of 
