Federal Register / Vol. 45, No. 227 / Friday, November 21, 1980 / Notices 
77397 
Experiments involving the use of plant 
genomes to propagate DNA sequences 
from eukaryotic viruses will be 
evaluated by NIH on a case-by-case 
basis (45) and will be conducted under 
the prescribed physical and biological 
containment conditions. (See Section 
IV-E-l-b-(3)-(c).) 
Ill— C— 4. Plant Host-Vector Systems 
Other than Viruses. (48) Organelle, 
plasmid, and chromosomal DNAs may 
be used as vectors. DNA recombinants 
formed between such vectors and host 
DNA, when propagated only in that host 
(or a closely related strain of the same 
species), are exempt from these 
guidelines (see Section I-E). DNA 
recombinants formed between such 
vectors and DNA from cells other than 
the host species require P2 physical 
containment. The development of host- 
vector systems that exhibit a high level 
of biological containment, such as those 
using protoplasts or undifferentiated 
cells in culture, permit (2A) a decrease 
in the physical containment to PI. 
Intact plants or propagative plant 
parts which cannot be grown in a 
standard P2 laboratory because of their 
large size may be grown under the Pi 
conditions described above in Section 
III— C— 3, except that (i) sterilization of 
run-off water is required where this is a 
plausible route for secondary infection 
and (ii) the standard P2 practices are 
adopted for microbiological work, and 
(iii) negative air pressure should be 
employed in the greenhouse or growth 
chamber when infectious agents are 
used which generate airborne 
propagules. 
Ill— C— 5. Fungal or Similar Lower 
Eukaryotic Host- Vector Systems. 
Certain certified HVl and HV2 host- 
vector system appear in Appendix D. 
The containment levels for these 
systems are given in the subsections of 
Section III-A. Other systems in the 
future may be certified as HVl and HV2. 
At the time of certification, they may be 
added to Appendix D (and thus the 
containment levels for their use will be 
those of the subsections of Section III— 
A). Alternatively, at the time of their 
certification, another classification of 
containment levels for experiments 
using them may be assigned by NIH. 
In addition to the experiments 
described above, the following 
experiments may be carried out without 
the eukaryotic host (Host C) having 
been approved as an HVl host: DNA 
from Host C may be inserted into a 
vector and propagated in E. coli K-12 
under Pi conditions. Subsequently, this 
recombinant DNA may be returned to 
Host C and propagated there under PI 
conditions. (43) 
Containment levels for other classes 
of experiments involving non-HVl. 
systems may be expressly approved by 
the Director, NIH. (See Sections IV-E-1- 
b— (1)— (b). IV-E-l-b-(2)-(c), and IV-E-1- 
b— (3)— (b).) 
Ill— 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 vector and propagated 
in E. coli K-12 under Pi containment 
conditions. 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) 
III— C— 7. Transfer of cloned DNA 
Segments to Eukaryotic Organisms. 
III-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 PI 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 
(2A). Transfers to any other host will be 
considered by the RAC on a case-by- 
case basis (45). 
III-C-7-b. Transfer to Higher Plants. 
DNA from any nonprohibited source 
[Section I-D] which has been cloned 
and propagated in E. coli or S. 
cerevisiae under Pi conditions, may be 
transferred with the E. coli or S. 
cerevisiae vector used for cloning to any 
higher plant organisms (Angiosperms 
and Gymnosperms) and progagated 
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 undr Pi conditions 
described undr Section III— C— 3. 
Containment must he 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 other plant organisms 
will be considered on a case-by-case 
basis [45). 
Ill— D. Complementary DNAs. Specific 
containment levels are given in Section 
III-A-2-a (see also last column of Table 
III) for complementary DNA (cDNA) of 
viral mRNA. For the other Sections of 
the Guidelines, where applicable, 
cDNAs synthesized in vitro are included 
within each of the above classifications. 
For example, cDNAs formed from 
cellular RNAs that are not purified and 
characterized are included under III— A— 
1, shotgun experiments; cDN’s formed 
from purified and characterized RNAs 
are included under III— A— 3: etc. 
Due to the possibility of nucleic acid 
contamination of enzyme preparations 
used in the preparation of cDNAs, the 
investigator must employ purified 
enzyme preparations that are free of 
viral nucleic acid. 
III— E. Synthetic DNAs. If the synthetic 
DNA segment is likely to [2A] yield a 
potentially harmful polynucleotide or 
polypeptide (e.g., a toxin or a 
pharmacologically active agent), the 
containment conditions must be as 
stringent as would be used for 
propagating the natural DNA 
counterpart. 
If the synthetic DNA sequence codes 
for a harmless product, (2A) it may be 
propagated at the same containment 
level as its purified natural DNA 
counterpart. For example, a synthetic 
DNA segment which corresponds to a 
nonharmful gene of birds, to be 
propagated in Saccharomyces 
cerevisiae, would require P2 physical 
containment plus an HVl host-vector, or 
PH- HV2. 
If the synthetic DNA segment is not 
expressed in vivo as a polynucleotide or 
polypeptide product, the organisms 
containing the recombinant DNA 
molecule are exempt [4) from the 
Guidelines. 
IV. Roles and Responsibilities 
IV- A. Policy. Safety in activities 
involving recombinant DNA depends on 
the individual conducting them. The 
Guidelines cannot anticipate every 
possible situation. Motivation and good 
judgement are the key es~~. ‘ials to 
protection of health and 
environment. 
The Guidelines are intended to help 
the Institution, the Institutional 
Biosafety Committee (IBC), the 
Biological Safety Officer, and the 
Principal Investigator determine the 
safeguards that should be implemented. 
These Guidelines will never be complete 
or final, since all conceivable 
experiments involving recombinant 
DNA cannot be foreseen. Therefore, it is 
the responsibility of the Institution and ' 
those associated with it to adhere to the 
purpose of the Guidelines as well as to 
their speifics. 
Each Institution (and the IBC acting 
on its behalf) is responsible for ensuring 
that recombinant DNA activities comply 
with the Guidelines. General recognition 
of institutional authority and 
responsibility properly establishes 
accountability for safe conduct of the 
research at the local level. 
[16] 
