Figure 5.

Efficient Transduction of Bicistronic Fluorescent Reporter Vectors Using AAV1-PHP.B

(A) Inserting a 7-mer PHP.B peptide sequence in between S588 and T589 of the AAV1 capsid to generate AAV1-PHP.B. (B) pCMV-iRFP720-GFP construct fusing iRFP720 infrared to the GFP fluorescent transgenes via the T2a bicistronic element. CMV promoter is used to drive the expression of bicistronic fluorescent reporter vectors. (C) AAV1-WT and AAV1-PHP.B transductions of HEK293FT cells. (D) AAV1-WT and AAV1-PHP.B transductions of U87 cells. After 24 h of incubation with 0.1× ViraDuctin, AAVs were added to the U87 cells. GFP fluorescent images were taken 3 days after the first transduction (for single transduction) or 2 days after the second transduction (for double transduction). Second transduction was performed 24 h after the first transduction. In the flow cytometry analysis (right), the fluorescent channels APC-H7 and FITC were used to detect the iRFP720- and GFP-expressing cells, respectively. The percentage of cells expressing both iRFP720 and GFP are indicated in the density plot.