Appendix 1 
Recombinant Adenovirus 
ACN53 
^lajorJjateTranscri^^ 
.HI 
E 3 Deletio n 
Ad5<fl327 
Delete Ela,Elb and pDC 
pK 
ITR CMV 
TPL 
P 53 
Figure 1 This schematic diagram depicts the genomic structure of ACN53. The p53 
expression cassette replaces the endogenous Ad5/dl327 Early region 1 (El). The locations 
of early regions 1 through 4 (E1-E4), protein IX gene (pDC), inverted terminal repeat 
(UK), human cytomegalovirus imm ediate early promoter/enhancer (CMV), adenovirus 
type 2 tripartite leader (TPL) and p53 cDNA are indicated. 
ACN53 is similar to other recombinant adenoviral constructs reviewed by the RAC and FDA, 
except that it contains the additional deletion of the pDC coding sequence. This deletion reduces the 
homology of the viral DNA with that of the adenoviral sequences in the human 293 cell line, used 
for the production of ACN53. Therefore, it is expected that this deletion will result in a lower 
frequency of replication competent adenovirus arising during virus production than observed with 
viruses deleted for only El a and Elb. In addition, this deletion reduces the packaging capacity of 
the virus by approximately 3 Kb, such that an ACN53 virus genome that may have acquired 
additional genetic information from the 293 cells cannot be packaged efficiendy. 
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Recombinant DNA Research, Volume 20 
