The hyperactive Sleeping Beauty transposase SB100X improves the genetic modification of T cells to express a chimeric antigen receptor

Z. Jin, S. Maiti, H. Huls, H. Singh, S. Olivares, L. Mátés, Z. Izsvák, Z. Ivics, D. A. Lee, R. E. Champlin, L. J.N. Cooper

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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 languageEnglish
Pages (from-to)849-856
Number of pages8
JournalGene Therapy
Volume18
Issue number9
DOIs
Publication statusPublished - Sep 1 2011

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Keywords

  • CD19
  • SB100X
  • SB11
  • Sleeping Beauty
  • T cells
  • chimeric antigen receptor

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

Cite this

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. (2011). The hyperactive Sleeping Beauty transposase SB100X improves the genetic modification of T cells to express a chimeric antigen receptor. Gene Therapy, 18(9), 849-856. https://doi.org/10.1038/gt.2011.40