Abstract
Sleeping Beauty (SB 3) 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 (CAR 3) 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.
Original language | English (US) |
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Pages (from-to) | 849-856 |
Number of pages | 8 |
Journal | Gene Therapy |
Volume | 18 |
Issue number | 9 |
DOIs | |
State | Published - Sep 2011 |
Keywords
- CD19
- SB100X
- SB11
- Sleeping Beauty
- T cells
- chimeric antigen receptor
ASJC Scopus subject areas
- Molecular Medicine
- Molecular Biology
- Genetics
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