1. Supplementary Figure 1 Wild-type HI-loop sequence :
5’-GAC ACA ACT
D T T
P S A
CCA AGT GCA-3’
Random oligonucleotide insertions:
Csp45I
Randomized oligonucleotide
SpeI
5’-GAC ACA ACT TTC GAA NNK NNK NNK NNK NNK NNK NNK ACT AGT CCA AGT GCA-3’ D T T F E X X X X X X X T S P S A
Oligonucleotide (SYENFSA) insertion:
5’-GAC ACA ACT TTC GAA TCG TAT GAG AAT TTT AGT GCG ACT AGT CCA AGT GCA-3’
Csp45I
SpeI D T F E S Y N A P
SYENFSA
Oligonucleotide (IVRGRVF) insertion:
5’-GAC ACA ACT TTC GAA ATT GTT CGT GGT CGG GTG TTT ACTAGT CCA AGT GCA-3’
Csp45I
SpeI D T F E I V R G S P A
IVRGRVF
Modified HI-loop sequences in adenovirus genome. In the adenovirus library, 21bp of random oligonucleotides were inserted between Csp45I and SpeI sites in the HI-loop of fiber coding region. In AdDCAR-SYE and AdDCAR-IVR, oligonucleotides (TCGTATGAGAATTTTAGTGCG: SYENFSA, ATTGTTCGTGGTCGGGTGTTT: IVRGRVF) were inserted, respectively.

2. Supplementary Figure 2 PC3 AsPC-1 Relative adenovirus DNA 30 Day 3 6025 50
Day 9
Day 7 20 40
Relative adenovirus DNA 15 30 10 20 5 10
AdDCAR
AdDCAR
-SYE
Ad-EGFP
AdDCAR
AdDCAR
-SYE
Ad-EGFP
Replication of adenovirus DNA in infected cells. PC3 or AsPC-1 cells were infected with AdDCAR, AdDCAR-SYE and Ad-EGFP at MOI of 30. Cells were harvested at each day and genomic DNA was extracted for quantitative PCR analysis of fiber knob and cellular GAPDH (730 Real Time PCR System: Applied Biosystems, Foster City, CA). The amount of capsid DNA in each sample was normalized to GAPDH at each time point, and compared with adenovirus DNA at day 3.