Federal Register / 
standard P2 laboratory because of their 
large size may be grown under the Pi 
I conditions describe above in Section 
Ul-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. 
lU-C-S. Fungal or Similar Lower 
Eukaryotic Host- Vector Systems. 
Certain certified HVl and HV2 host- 
vector systems appear in Appendix D. 
The containment levels for these 
systems are given in the subsections of 
Section Ill-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. 
Subsequently, this recombinant DNA 
may be returned to Host C and 
propagated there under Pi conditions. 
(4J) Containments levels for other 
classes of experiments involving non- 
HVl systems may be expressly 
approved by the Director, NIH. (see 
Sections IV-E-l-b-(l>-0)). IV-E-l-b- 
(2Hc). and IV-E-l-lH3Hb).) 
Ill-C-e. 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. 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) 
ni-C-7. Transfer of Cloned DNA 
Segments to Eukaryotic Organisms. 
IU-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 doned and propagated in E. coli 
K-12, 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 
Vol. 46. No. 126 / Wednesday, July 1, 1981 / Notices 34475 
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). 
llI-C-7-b. Transfer to Higher Plants. 
DNA from any nonprohibited source 
(Section I-D] which has been cloned 
and propagated in E. coli K-12 or S. 
cerevisiae, may be transferred with the 
E. coli or S. cerevisiae 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 IU-C-3. 
Containment must be modified to ensure 
that the spread of pollen, seed or other 
propagules is prevented. This can be 
accompanied by conversion to negative 
pressure in the growth cabinet or 
greenhouse or by physical entrapment 
by “bagging” or reproductive structures. 
Transfers to any other plant organisms 
will be considered on a case-by-case 
basis (45). 
IIl-D. Complementary DNAs. Specific 
containment levels are given in Section 
IU-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 lU-A- 
1. shotgun experiments; cDNAs formed 
from puriHed and characterized RNAs 
are included under lIl-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 nudeic 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 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 
nopharmful gene of birds, to be 
propagated in saccharomyces 
cerevisiae, would require P2 physical 
containment plus an HVl host-vector, or 
Pl-t-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 essentials to 
protection of health and the 
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 specifics. 
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. 
The following roles and 
responsibilities constitute an 
administrative framework in which 
safety is an essential and integral part of 
research involving recombinant DNA 
molecules. Further clarifications and 
interpretations of roles and 
responsibilities will be issued by NIH as 
necessary. 
IV-B. General Applicability. The 
Guidelines are applicable to all 
recombinant DNA research within the 
United States or its territories which is 
conducted at or sponsored by an 
Institution that receives any support for 
recombinant DNA research from NIH. 
This includes research performed by 
NIH directly. 
An individual receiving support for 
research involving recombinant DNA 
must be associated with or sponsored 
by an Institution that can and does 
assume the responsibilities assigned in 
these Guidelines. 
The Guidelines are also applicable to 
projects done abroad if they are 
supported by NIH funds. If the host 
country, however, has established rules 
for the conduct of recombinant DNA 
projects, then a certificate of compliance 
[ 163 ] 
