33120 
NOTICES 
vu attended by 27 distinguished virol- 
ogists and other scientists from the 
United States, the United Kingdom. 
West Germany. Finland, Prance. 
Sweden, and Switzerland. A report was 
published In the Federal Register on 
March 31, 1978 (43 FR 13748) and con- 
stitutes appendix E to the present doc- 
ument. The "Ascot Workshop" con- 
cluded: 
The probability that K13 organism* carry- 
las viral DNA Inserts could represent a sig- 
nificant hazard to the co mmuni ty la so 
small as to be of no practical consequence 
• • • . Viral genomes or fragments thereof, 
cloned In K. raft K13 using approved plas- 
mid or phage vectors pose no more risk than 
work with the Infectloua virus or Its nucleic 
add and In most, if not all ca s es , clearly 
present lees risk. In fact, the workshop par- 
ticipants agreed that cloning of viral DNA 
In E. coil K12 may provide a unique oppor- 
tunity to study with greatly reduced risks 
the biology of extremely pathogenic and vir- 
ulent druses. 
On April 8-7, 1978 (ag announced In 
the Mar. 17 Federal Register), an 
RAC-sponsored working group com- 
posed of distinguished American mi- 
crobiologists met to review the report 
of the Ascot Workshop. The report of 
this working group Is appendix F. The 
group unanimously endorsed the 
Ascot report with certain minor 
amendments. Their report was pre- 
sented to the RAC. was unanimously 
accepted, and has been substantially 
adopted In part 111 of the PKONTH. 
Characterized, and Pm rifled Clones. 
Concern was expressed by several com- 
mentators about the revisions In the 
PRO-RAC which would allow the 
local IBC (with notification to be sent 
to NIH) to reduce either the biological 
or physical containment level by one 
step If (1) the DNA ts 99-percent puri- 
fied and shown to be free of harmful 
genes before Its Insertion Into a recom- 
binant molecules or 12) the clone repli- 
cating the DNA la rigorously charac- 
terized and free of harmful genes. In 
the original guidelines, the redaction 
In case (2) could only be done with 
prior NIH approval. 
There was support from several com- 
mentators for the changes In this sub- 
section. The rationale Is explained In 
the PRO-RAC and the PRO-NIH: 
Many of the risks which m i g h t ranee lv- 
ably arise from some ty p— of recemtotaant 
DBA expe rim ents, particularly shotgun ex- 
periments, would result frost the toadver- 
tant cloning of a harmful sequence. There- 
for*. In caaea where the risk of Inadvertent- 
ly Cloning the "wrong" DNA Is reduced by 
prior enrichment for the desi r e d piece, or In 
which a done, made from a r and o m assort- 
ment of DNA’a has been p ur i f ied and the 
ahaence of harmful sequence* eetabhehed. 
the cents lamen t- conditions for farther 
work may be reduced. 
Some commentator* noted the ambi- 
guity and difficulty attendant ta the 
Phraoe free of harmful genes.'* The 
aforemei.:foned EMBO Co mm ft tec re- 
ports that "several national guidelines 
for recombinant DNA r e s e a rch state 
that containment measures may be re- 
laxed once a cloned DNA fragment 
has been biochemically characterized 
and shown to be free of harmful genes 
(NIH guidelines) or devoid of any 
known pathogenic characteristic 
(French guidelines). The EMBO Com- 
mittee believes the latter to be a more 
feasible requirement, but neither can 
readily be met. and the committee 
finds It difficult to suggest what sorts 
of experimental tests might be devised 
to meet these requirements." 
The terms "characterized and “free 
of harmful genes" are unavoidably 
vague. However, footnote 3 of the 
PRO-NIH goes on to list five types of 
data which should be considered In 
making this determination. 
Some commentators were also con- 
cerned that this granting of additional 
authority to the local IBCs for single- 
step lowering In containment levels 
might Introduce variability In the ap- 
plication of the guidelines. NIH. 
having considered that possibility, has 
decided that the principle of promot- 
ing local involvement In the tmplemen-. 
tatlon of the guidelines outweighs the 
difficulties that may be enco unt er e d 
In this process. In an effort to mini 
mfcse theae problems, NIH has (I) at- 
tempted to make all parts of the 
guide lin es as clear, specific, and unam- 
biguous as possible and (2) expanded 
the “roles and responsibilities" section 
to outline functions and responsibil- 
ities in greater detail. 
Also, the guidelines require that the 
Office of Recombinant DNA Activities 
at the NTH be notified In writing of 
such an action. A mechanism Is there- 
fore in place to Insure that such ac- 
tions proceed with an acceptable 
degree of uniform! ty. 
The question was raised whether a 
clone of which the containment level 
was lowered by the IBC at Institution 
X may. after ship m ent to fnatftwtton 
T. be used at the lower level without 
review by T*a IBC. It has clearly been 
the hrtentlon of both the RAC and 
NIH that the IBC at the receivi ng tn- 
stltutfon must approve the reduction 
in conta inment for the handling of the 
clone hr such a situation. The I nycst l- 
gatorat the receiving Institution, how- 
ever, must handle the done at the 
higher level until such permission is 
grant ed . 
One commentator urged that prior 
ckmtnr be accepted as a technique far 
the purification of DNA mo le cules 
before their reinsertion Into a r e cotii - 
btnent mole cu le. The PRO-RAC speci- 
fied that purification must be 
ac h ieved “by physical or c h e mica l 
teehnlqnea“ The criterion fbr the 
single-step reduction hr containment 
le v e W In this situation Is that the DNA 
preparation be 99-percent pure. There 
Is no reason, the commentator held, to 
restrict the means by which such puri- 
fication Is attained. This suggestion 
has been accepted. The words "by 
physical and chemical techniques" fol- 
lowing the work "purified" have been 
stricken from the FRG-NIH, better 
serving the needs of the investigator 
without reducing the margin of safety 
to the public and the environment. 
One commentator noted that the 
PRO-RAC might be Interpreted aa al- 
lowing a single-step reduction in con- 
tainment levels tor purification of the 
DNA before Its Insertion Into a recom- 
binant molecule, and a further stngle- 
step reduction In conta i n m e n t once 
the same m olecule has been cloned. 
This was not Intended. Therefore, 
clarifying language has been added In 
the PRO-NIH stating that an IBC 
"may give approval for a single-step 
reduction bi physical or biological con- 
tainment on receipt of evidence of 
characterisation ef a clone derived 
from a shotpm experiment 9 • •* 
Permlssable Experiments With Eukar- 
yotic Hast- Vectors 
Viral Vectors. A number of commen- 
tators felt that the stringent contain- 
ment conditions required, both in the 
original guidelines and ha the PRO- 
RAC. for Introduction of recombinant 
DNA Into tissue culture cells, using 
viruses as vectors, were unwarranted. 
Tbs EMBO Standing Advisory Com- 
mittee on Recombinant DNA Re- 
search wrote; 
la experiments Inrolvlns the Introduction 
o i foreign DNA Into cultured cells o t ani- 
mals using DNA viruses as vectors, biologi- 
cal containment la assured by the very re- 
stricted permissive conditions for the host 
cells; the only routes by which the recom- 
binant m decide might es ca p e ore by chance 
Infection at s contaminating microorganism 
or within a viral rspslrt and the rise at the 
recombinant molecule may weO preclude 1th 
encapsulation * * *. F* example, cloning of 
mouse DNA using polyoma virus as a veetor 
and mouse cells as host should not require 
precautions more stringer it than those rou- 
tinely need for many years hr laboratories 
stadyhig. polyoma virus Infection at amuse 
cads mid alca The EMBO Committee finds 
the proposals tem thfe dasa at experiments 
in the revised NIB Guideline* not suffi- 
ciently discriminating because they would 
Impose unnecessarily high levels of physical 
conta inm ent fbr experiments with marry eu- 
karyotic DWAV. 
Discussed fariler wtthfn the present 
document was the ascot Workshop 
report (appendix El and the report of 
the Working Group that met on April 
8-T, 1FTR (Appendix FV The recam- 
me nds tluns of the Working* OTOup 
have bdetr ac cepte d and I nc o rpor ated 
ftrto the FRG-NIH. 
Plant lEast-Tector Systems, pla- 
c u s Bril earlier was the W brtshdp oh 
Rfc* Assessment of Agricultural Fsth- 
otetts, heftf on ‘ March fbj-ft. 1978. 
under the- sponsor sh ip of U'SDA, NSf. 
ndcral Boom, roe. a n* isa— me 
[ 81 ] 
mt, Mir jr mt 
