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

Research output: Contribution to journalArticle

60 Citations (Scopus)

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 2011

Fingerprint

Transposases
Beauty
Antigen Receptors
T-Lymphocytes
Plasmids
CD19 Antigens
Cell Engineering
Founder Effect
Introduced Species
Genetic Engineering
Human Engineering
DNA
Medical Genetics
Antigen-Presenting Cells
Genome
Messenger RNA

Keywords

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

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

Cite this

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

In: Gene Therapy, Vol. 18, No. 9, 09.2011, p. 849-856.

Research output: Contribution to journalArticle

Jin, Z, Maiti, S, Huls, H, Singh, H, Olivares, S, Mátés, L, Izsvák, Z, Ivics, Z, Lee, DA, Champlin, RE & Cooper, LJN 2011, 'The hyperactive Sleeping Beauty transposase SB100X improves the genetic modification of T cells to express a chimeric antigen receptor', Gene Therapy, vol. 18, no. 9, pp. 849-856. https://doi.org/10.1038/gt.2011.40
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. / The hyperactive Sleeping Beauty transposase SB100X improves the genetic modification of T cells to express a chimeric antigen receptor. In: Gene Therapy. 2011 ; Vol. 18, No. 9. pp. 849-856.
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