Attachment II - Pg. 5 
2. Research methods 
Provide a full description of the methods and reagents to be employed for 
gene delivery, the rationale for their use, and the experimental basis (de- 
rived from tests in cultured cells and experimental animals) for claims 
about efficacy and safety. Include precise means to evaluate the success of 
therapy with laboratory methods; an assessment of the risks of infectious 
agents in the recipients; and an evaluation of the likelihood of Inducing 
additional disease or Invading the germ line. The folloving are points to 
be addressed. 
a. What is the structure of the cloned ONA that will be used? 
(1) Describe the gene (genomic or cDNA), the bacterial plasmid or phage 
vector, and the delivery vector (If any). Provide complete sequence 
analysis or a detailed restriction map of the total construct. 
(2) What regulatory .elements does the construct contain (e.g., promo- 
ters, enhancers, polyadenylatlon sites, replication origins, etc.)? 
(3) Describe the steps used to derive the DNA construct. 
b. What is the structure of the material that will be given to the pa- 
tient? 
(1) Describe the preparation and structure of all materials that will 
either be given to the patient or used to treat the patient's 
cel Is. 
(a) If DNA, what is the purity (both In terms of being a single DNA 
species and In terms of other contaminants)? What tests were 
used and what Is the sensitivity of the tests? 
(b) If a virus, what cell is it grown In (any special features)? 
What medium and serum are used? How is the virus purified? 
What is its structure and purity? What steps are being taken 
(and assays used with their sensitivity) to detect and eliminate 
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