Fig. 7: piggyBat transposition in cultured human cells and the effects of mutating predicted N-terminal casein kinase II phosphorylation sites and duplicating the CRD.

a pBat (top) and pB (bottom) N-terminal amino acid sequences highlighting the CKII phosphorylation motifs (underlined). The numbering above corresponds to the amino acid number of pBat. b Transposition activity for WT and phosphorylation mutant transposases. Data are presented as mean values +/− SD. Data for point mutants (n = 4 biological replicates) were compared to WT (n = 8) using two-tailed unpaired t tests. ****, p < 0.0001. The experiment has been performed twice with similar results. c Transposition activity for WT and 4StoA point mutant transposase on truncated ends (n = 4 biological replicates). Data are presented as mean values +/− SD. P-values were determined by a two-tailed unpaired t test. **, p = 0.002. The experiment has been performed twice with similar results. d Transposition activity of WT and pBat 4StoA compared to pB Δ74 2xCRD on piggyBac LE35/LE35. Data are presented as mean values +/− SD. P-values were determined by a two-tailed unpaired t test. ****, p = 0.0001. n = 4 biological replicates. The experiment has been performed twice with similar results. e Schematic representation of pBat-4StoA-2xCRDV1. Created in BioRender. Hickman, A. (2024) https://BioRender.com/v39k454. f Comparison of transposition activity of pBat-4StoA, pBat-4StoA-2xCRDV1, and pBat-4StoA-2xCRDV2 on truncated piggyBat transposon ends (n = 4 biological replicates); **, p = 0.0014. Data are presented as mean values +/− SD. The experiment has been performed twice with similar results. g Schematic representation of pBat-4StoA-2xCRDV2. Created in BioRender. Hickman, A. (2024) https://BioRender.com/v39k454. h Transposition activity of pBat-4StoA-2xCRDV2 on truncated piggyBat transposon ends (n = 2 biological replicates). The experiment has been performed twice with similar results. Source data are provided as a Source Data file.