Figure 4.

Characterization of Velcro-AAV interaction with complementary LZ peptides. Velcro-AAV E3 (A) and K3 (B) vectors were incubated with complementary His₆ tagged peptides (300 000×, 30 000×, 3000×, or 300×), then the mixture was loaded onto nickel columns, and the fraction of total virus eluted in the flow through (S), wash (W), and elution fractions (E1, E2, E3, E4) was quantified via qPCR. Total virus in elution fractions was summed; the virus in individual fractions is plotted in Figure S3. N = 3 independent experiments, error bars are SEM. Two-way ANOVA was performed with Dunnett’s posthoc multiple comparison test to compare mutants to AAV9 with 300 000× peptide in each wash fraction. **p < 0.01, ***p < 0.001. C) Velcro-AAV were incubated with complementary His₆ tagged peptides at 300 000× and then used to transduce CHO-Lec2 cells at an MOI of 5000. Transgene expression was measured by flow cytometry at 48 h post-transduction. Transduction index (the product of %+GFP cells and geometric mean fluorescence intensity) is reported relative to AAV9 with no peptide. N = 3 independent experiments; error bars are SEM. One-way ANOVA was performed with Sidak’s posthoc multiple comparison test to compare all experimental conditions to AAV9 with no added peptide and to compare each capsid–peptide transduction to the corresponding capsid–no peptide control. ***p < 0.001.