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- Title
The hyperactive Sleeping Beauty transposase SB100X improves the genetic modification of T cells to express a chimeric antigen receptor.
- Authors
Jin, Z; Maiti, S; Huls, H; Singh, H; Olivares, S; Mátés, L; Izsvák, Z; Ivics, Z; Lee, D A; Champlin, R E; Cooper, L J N
- Abstract
Sleeping Beauty (SB3) transposon and transposase constitute a DNA plasmid system used for therapeutic human cell genetic engineering. Here we report a comparison of SB100X, a newly developed hyperactive SB transposase, to a previous generation SB11 transposase to achieve stable expression of a CD19-specific chimeric antigen receptor (CAR3) in primary human T cells. The electro-transfer of SB100X expressed from a DNA plasmid or as an introduced mRNA species had superior transposase activity in T cells based on the measurement of excision circles released after transposition and emergence of CAR expression on T cells selectively propagated upon CD19+ artificial antigen-presenting cells. Given that T cells modified with SB100X and SB11 integrate on average one copy of the CAR transposon in each T-cell genome, the improved transposition mediated by SB100X apparently leads to an augmented founder effect of electroporated T cells with durable integration of CAR. In aggregate, SB100X improves SB transposition in primary human T cells and can be titrated with an SB transposon plasmid to improve the generation of CD19-specific CAR+ T cells.
- Subjects
TRANSPOSONS; T cells; GENETIC engineering; GENE expression; ANTIGENS; PLASMIDS; DNA; CD antigens
- Publication
Gene Therapy, 2011, Vol 18, Issue 9, p849
- ISSN
0969-7128
- Publication type
Article
- DOI
10.1038/gt.2011.40