Recombinant DMA Advisory Committee - 3/3-4/94 
for protocol approval. A large segment of vector sequence not accountable is not 
acceptable. 
Ms. Grossman emphasized that for approval a vector must be completely sequenced or a 
detailed restriction enzyme map provided. A Southern blot analysis of cells transduced 
with the antisense K-ras and p53 retroviral constructs, and a comparison with the LNSX 
vector control will be required. In addition, the Northern blot data on vector transcript 
should be provided. Dr. Roth said that Northern blot analysis will be difficult for tumor 
RNA. 
Dr. Motulsky stated that although the animal data is impressive, a full characterization of 
the vectors is essential for this protocol. 
In terms of characterization of the vector structures regarding both retroviral constructs 
containing the antisense K-ras and p53 genes, Drs. Post, Straus, and Ms. Grossman stated 
that the following information is necessary: (1) the complete DNA sequences of the 
vector constructs, (2) a detailed restriction enzyme digestion map of the constructs, and 
(3) a series of Southern blot analyses of vector DNA from cells transduced with the 
parental vectors versus constructs containing the gene inserts. Data should be obtained 
from several transduced cell lines demonstrating concordance with the restriction enzyme 
analysis and the restriction map. 
Committee Motion 
A motion was made by Dr. Straus and seconded by Dr. Parkman to approve the 
protocol. Approval of the protocol is contingent on the review and approval of the 
following data for both the K-ras and p53 retroviral constructs by the primary reviewers 
and Drs. Post and Straus: (1) complete vector sequences, including detailed restriction 
enzyme maps relevant to the LNSX backbone and the gene inserts; and (2) Southern 
blot analyses using several transduced cell lines, including cell lines transduced with the 
vector constructs with and without gene inserts, demonstrating concordance with the 
restriction enzyme 2 malysis and sequence data. The motion to approve the protocol 
passed by a vote of 13 in favor, 0 opposed, and no abstentions. 
XVI. CHAIR REPORT - OUTGOING MEMBERS 
Dr. Walters noted that several members of the RAC have completed their term of 
service. He thanked Drs. Carmen, Hirano, Post, Geiduschek and Krogstad, and Ms. 
Grossman, for their dedication, expertise, and tireless efforts, which have contributed 
significantly to the advancement of human gene therapy. 
XVII. REPORT FROM THE WORKING GROUP ON INFORMED CONSENT - 
AMENDMENTS TO PART I-D OF THE POINTS TO CONSIDER/DR, ZALLEN 
Dr. Gary Ellis, Director of the NIH Office for Protection from Research Risks, 
recommended several avenues that should be pursued by the RAC with regard to the 
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