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| Item Type: | Article |
|---|---|
| Title: | The hyperactive Sleeping Beauty transposase SB100X improves the genetic modification of T cells to express a chimeric antigen receptor |
| Creators Name: | Jin, Z., Maiti, S., Huls, H., Singh, H., Olivares, S., Mates, L., Izsvak, Z., Ivics, Z., Lee, D.A., Champlin, R.E. and Cooper, L.J. |
| 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. |
| Keywords: | Chimeric Antigen Receptor, T Cells, Sleeping Beauty, SB11, SB100X, CD19 |
| Source: | Gene Therapy |
| ISSN: | 0969-7128 |
| Publisher: | Nature Publishing Group |
| Volume: | 18 |
| Number: | 9 |
| Page Range: | 849-856 |
| Date: | September 2011 |
| Official Publication: | https://doi.org/10.1038/gt.2011.40 |
| PubMed: | View item in PubMed |
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