Transposition efficiency after infection with the transposon-donor vector AAV-neo co-transfected with transposase encoding plasmids.

<div><p>(<b>a</b>) Colony forming assay to determine integration efficiencies from the AAV-neo vector co-transfected with transposase encoding plasmids. HeLa-cells were first transfected with 1µg of the respective transposase encoding plasmid (pCMV-mSB, pCMV-HSB5, or pCMV-SB100X) and one day post-transfection cells were infected with AAV-neo at MOI 10,000. Two days post-infection cells were diluted and kept under selection pressure for 14 days. Obtained cell colonies were either collected as cell pools for integration site analysis or stained with methylene blue to determine transposition rates. Transposase encoding plasmids contain expression cassettes for the hyperactive transposases HSB5 and SB100X and the inactive SB transposase version mSB expressed under the control of the cytomegalovirus promoter (CMV).</p> <p>(<b>b</b>) Result of the colony forming assay. The Y-axis shows the number of neomycin resistant colonies obtained from 4 x 10⁵ cells in different experimental settings. The lower panel shows examples of original tissue culture plates from the different groups after methylene blue staining. Error bars indicate standard deviation (n=3). *Significant difference between the hyperactive transposase groups (SB100X and HSB5) and the mSB control group (p-value < 0.05).</p>