Abstract 3495: A nucleolus-predominant piggyBac transposase increases transposition efficiency in human cancer cells

Proceedings: AACR Annual Meeting 2014; April 5-9, 2014; San Diego, CA PiggyBac is a prevalent transposon system used to deliver transgenes into and to functionally explore the mammalian untouched genomic territory. Efforts to increase its transposition efficiency in mammals through engineering the corresponding transposase (PBase) codon usage to improve its expression level is reported. The hyperactive PBase developed through screening for mutant variants are also established. To increase the safety for its potential use in regenerative medicine applications, site-specific transposition using engineered Zn finger-, Gal4- fused PBase, and an excision-prone PBase variant are also successfully developed. Here we report the construction of a nucleolus predominant PBase, NP-mPB, by adding a nucleolus predominant (NP) signal peptide from HIV-1 TAT protein to a mammalian codon optimized PBase (mPB). Although there is a predominant fraction of the NP-mPB-tGFP fusion proteins concentrated in the nucleoli, an insertion site preference toward nuleolar organizer regions (NORs) is not detected. Instead a 3-fold increase in piggyBac transposition efficiency is found in human cancer cell line. Citation Format: Jin-Bon Hong, Fu-Ju Chou, Amy T. Ku, Hsiang-Hsuan Fan, Tung-Lung Lee, Yung-Hsin Huang, I-Chang Su, You-Tzung Chen. A nucleolus-predominant piggyBac transposase increases transposition efficiency in human cancer cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3495. doi:10.1158/1538-7445.AM2014-3495