title: The hyperactive Sleeping Beauty transposase SB100X improves the genetic modification of T cells to express a chimeric antigen receptor.
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.
authors: Division of Pediatrics, Children's Cancer Hospital, The University of Texas Graduate School of Biomedical Sciences, The University of Texas MD Anderson Cancer Center, Houston, USA.
authors: Z
authors: Z
authors: Jin
authors: S
authors: S
authors: Maiti
authors: H
authors: H
authors: Huls
authors: H
authors: H
authors: Singh
authors: S
authors: S
authors: Olivares
authors: L
authors: L
authors: Mátés
authors: Z
authors: Z
authors: Izsvák
authors: Z
authors: Z
authors: Ivics
authors: D A
authors: DA
authors: Lee
authors: R E
authors: RE
authors: Champlin
authors: L J N
authors: LJ
authors: Cooper
mesh_terms: D018941:Antigens, CD19 / Q000378:metabolism*; D045744:Cell Line, Tumor; D003602:Cytotoxicity, Immunologic; D018274:Electroporation; D018014:Gene Transfer Techniques; D006801:Humans; D009369:Neoplasms / Q000276:immunology; D012333:RNA, Messenger; D011946:Receptors, Antigen / Q000235:genetics / Q000378:metabolism*; D013601:T-Lymphocytes / Q000378:metabolism*; D019895:Transposases / Q000235:genetics*
chemical_list: D018941:Antigens, CD19; D012333:RNA, Messenger; D011946:Receptors, Antigen; D019895:Transposases; C494471:sleeping beauty transposase, human