Stable gene transfer and expression in human primary T cells by the Sleeping Beauty transposon system.
The Sleeping Beauty (SB) transposon system is a nonviral DNA delivery system in which a transposase directs integration of an SB transposon into TA-dinucleotide sites in the genome. To determine whether the SB transposon system can mediate stable gene expression in human T cells, primary peripheral blood lymphocytes (PBLs) were nucleofected with SB vectors carrying a DsRed reporter gene. Plasmids containing the SB transposase on the same molecule as (cis) or on a molecule separate from (trans) the SB transposon mediated long-term and stable reporter gene expression in human primary T cells. Sequencing of transposon:chromosome junctions confirmed that stable gene expression was due to SB-mediated transposition. In other studies, PBLs were successfully transfected using the SB transposon system and shown to stably express a fusion protein consisting of (1) a surface receptor useful for positive T-cell selection and (2) a "suicide" gene useful for elimination of transfected T cells after chemotherapy. This study is the first report demonstrating that the SB transposon system can mediate stable gene transfer in human primary PBLs, which may be advantageous for T-cell-based gene therapies.
Huang
Xin
X
Division of Blood and Marrow Transplantation, Department of Pediatrics, University of Minnesota Cancer Center, Minneapolis, MN 55455,USA.
Wilber
Andrew C
AC
Bao
Lei
L
Tuong
Dong
D
Tolar
Jakub
J
Orchard
Paul J
PJ
Levine
Bruce L
BL
June
Carl H
CH
McIvor
R Scott
RS
Blazar
Bruce R
BR
Zhou
Xianzheng
X
D000818:Animals; D004251:DNA Transposable Elements / Q000235:genetics*; D018014:Gene Transfer Techniques; D017930:Genes, Reporter / Q000502:physiology; D005822:Genetic Vectors; D006801:Humans; D008213:Lymphocyte Activation; D010957:Plasmids / Q000494:pharmacology*; D013552:Swine; D013601:T-Lymphocytes / Q000166:cytology / Q000378:metabolism; D014162:Transfection; D019076:Transgenes / Q000502:physiology*; D019895:Transposases / Q000235:genetics* / Q000378:metabolism*
D004251:DNA Transposable Elements; D019895:Transposases; C494471:sleeping beauty transposase, human