Figure 5.
Genome Editing in HSPCs by AAV6 and Clade F Vectors Requires Matched Nucleases
HSPCs were transduced with AAV6, AAV9, or AAV9-G505R vectors packaging (A–C and E) CCR5-PGK-GFP vector genomes or (D and E) AAVS1-SA-2A-GFP vector genomes. Samples were electroporated with CCR5- or AAVS1-specific ZFN mRNAs as indicated. GFP expression was measured at day 2 or day 10 post-electroporation by flow cytometry. (A) Graphs of day-2 GFP measurements from AAV-vector-only samples show rates of initial AAV transduction with CCR5-PGK-GFP vector genomes at MOIs of 5 × 104 or 5 × 105. (B) Cell viability was assessed by determining the frequency of events that fell within the live cell gate at day 2 by flow cytometry. (C and D) Flow cytometry analysis of stable GFP expression at day 10 with CCR5-PGK-GFP (C) or AAVS1-SA-2A-GFP (D) genomes. (E) Graphical representation of GFP expression using CCR5-PGK-GFP (left) or AAVS1-SA-2A-GFP (right) vectors at day 10.