NOTICES 
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III-C-4. Plant Host-Vector Systems 
Other than Viruses. Organelle, plas- 
mid. and chromosomal DNAs may be 
used • as vectors. DNA recombinants 
fprmed 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 re- 
combinants formed between such vec- 
tors 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 un- 
differentiated cells In culture, permit 
[2A1 a decrease in the physical con- 
tainment 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 (l) steril- 
ization of run-off water Is required 
where this is a plausible route for sec- 
ondary infection and (11) the standard 
1*2 practices are adopted for microbio- 
logical work. 
III-C-5. Fungal or Similar Lower 
Eukaryotic Host-Vector Systems. The 
containment criteria f or DNA recom- 
binant experiments using these host- 
vectors most closely resemble those 
for prokaryotes, rather than those for 
the preceding eukaryotes, since the 
host ceils usually exhibit a capacity 
for dissemination outside the labora- 
tory that 1s similar to that for bacte- 
ria. Therefore, the procedures estab- 
lished for certification of HV systems 
other than E. coli K-12 (Section II-D- 
2) will also apply to these fungal or 
similar lower eukaryotic host-vector 
systems. 
Once a HV1 system is approved by 
NIH. It may be used under P2 contain- 
ment for shotgun experiments with 
phages, plasmids, and DNA from Class 
1 prokaryotes! 1] and lower eukaryotes 
that do not produce polypeptide 
toxins. [34] Other classes of recombin- 
ant DNA experiments with these HV 1 
systems will require prior approval 
and classification by NIH. (See Sec- 
tions rV-E-l-b-UMb). IV-E-l-b-<2>- 
(c) and IV-E-l-b-(3Mb).) If HV2 or 
HV3 systems of this type are devel- 
oped and approved by NIH. guidelines 
for their use in other types of recom- 
binant DNA experiments will also be 
established. 
In addition to the experiments de- 
scribed above, the following experi- 
ments may be carried out without the 
eukaryotic host (Host C) having been 
approved as an HV1 host; DNA from 
Host C may be Inserted into a vector 
from a certified EK2- host-vector 
system and propagated in E. coli K-12 
under the appropriate containment 
conditions [see Section III-A-l-<a)- 
(5)). 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 sys- 
tems may be expressly approved b^ 
the Director. NIH. (See Sections IV-E- 
l-b-dHb), IV-E-l-b-<2Mc) and IV- 
E-l-b-OMb).) 
III-D. Complementary DNAs. Specif- 
ic containment levels are given in Sec- 
tion III-A-a (see also last column of 
Table III) for complementary DNA 
(cDNA) or viral mRNA. For the other 
Sections of the Guidelines, where ap- 
plicable, 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 in- 
cluded under III-A-1. shotgun experi- 
ments; cDNAs 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 syn- 
thetic DNA seqment is likely to [2A] 
yield a potentially harmful polynu- 
cleotide or polypeptide (e.g.. a toxin or 
a pharmacologically active agent), the 
containment conditions must be as 
stringent as would be used for propa- 
gating the natural DNA counterpart. 
If the synthetic DNA sequence codes 
for a harmless product. 12A] it may be 
propagated at the same containment 
level as its purified natural DNA coun- 
terpart. For example, a synthetic DNA 
segment which corresponds to a non- 
harmful gene of birds, to be propagat- 
ed in E. coli K-12. would require Ps 
physical containment plus an EK1 
host-vector, or PI + EK2. 
If the synthetic DNA segment is not 
expressed in vivo as a polynucleotide 
or polypeptide product, the organisms 
containing the recombinant DNA mol- 
ecules are exempt[4] from the Guide- 
lines. 
IV. Rules and Responsibilities 
IV- A. Policy. Safety in activities in- 
- volving 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 envi- 
ronment. 
The Guidelines are intended to help 
the Institution, the Institutional Bio- 
safety Committee (IBC), the Biologi- 
cal 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 responsi- 
bility of the Institution and those as- 
sociated with it to adhere to the pur- 
pose of the Guidelines as well as to 
their specifics. 
Each Institution (and the IBC acting 
on its behalf is responsible for ensur- 
ing 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 responsibil- 
ities constitute an administrative 
framework in which safety is an essen- 
tial and integral part of research in- 
volving recombinant DNA molecules. 
Further clarifications and interpreta- 
tions of roles and responsibilities will 
be issued by NIH as necessary. 
IV-B. General Applicability. The 
Guidelines are applicable to all recom- 
binant DNA research within the 
United States or its territories which 
is conducted at or sponsored by an In- 
stitution 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 with those rules may be 
submitted to NIH In lieu of compli- 
ance with NIH Guidelines. NIH re- 
serves the right to withhold funding if 
the safety practices to be employed 
abroad are not reasonably consistent 
with the NIH Guidelines. 
IV-C. General Definitions. The fol- 
lowing terms, which are used through- 
out the Guidelines, are defined as fol- 
lows; 
IV-C-1. "DNA" means deoxyribonu- 
cleic acid. 
IV-C-2. "Recombinant DNA" or "re- 
combinant DNA molecules'* means 
either (i) molecules which are con- 
structed outside living cells by Joining 
natural or synthetic DNA segments to 
DNA molecules that can replicate in a 
living cell, or (ii) DNA molecules 
which result from the replication of a 
molecule described in (i) above. 
IV-C-3. "Memorandum of Under- 
standing and Agreement” or "MUA” is 
a document that (1) provides to NIH or 
other Federal funding agency an Insti- 
tution's certification that the recom- 
binant DNA research project complies 
with the NIH Guidelines and (ii) con- 
tains other essential data as required 
FEOERAi REGISTER, VOL 41, NO. 247— FRIDAY, DECEMBER 22. 1971 
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