Gene transfer efficiency and genome-wide integration profiling of Sleeping Beauty, Tol2, and piggyBac transposons in human primary T cells.
In this study, we compared the genomic integration efficiencies and transposition site preferences of Sleeping Beauty (SB or SB11), Tol2, and piggyBac (PB) transposon systems in primary T cells derived from peripheral blood lymphocytes (PBL) and umbilical cord blood (UCB). We found that PB demonstrated the highest efficiency of stable gene transfer in PBL-derived T cells, whereas SB11 and Tol2 mediated intermediate and lowest efficiencies, respectively. Southern hybridization analysis demonstrated that PB generated the highest number of integrants when compared to SB and Tol2 in both PBL and UCB T cells. Tol2 and PB appeared more likely to promote clonal expansion than SB, which may be in part due to the dysregulated expression of cancer-related genes near the insertion sites. Genome-wide integration analysis demonstrated that SB, Tol2, and PB integrations occurred in all the chromosomes without preference. Additionally, Tol2 and PB integration sites were mainly localized near transcriptional start sites (TSSs), CpG islands and DNaseI hypersensitive sites, whereas SB integrations were randomly distributed. These results suggest that SB may be a preferential choice of the delivery vector in T cells due to its random integration site preference and relatively high efficiency, and support continuing development of SB-mediated T-cell phase I trials.
Huang
Xin
X
Division of Pediatric Blood and Marrow Transplantation, University of Minnesota Medical School, Minneapolis, Minnesota, USA.
Guo
Hongfeng
H
Tammana
Syam
S
Jung
Yong-Chul
YC
Mellgren
Emil
E
Bassi
Preetinder
P
Cao
Qing
Q
Tu
Zheng Jin
ZJ
Kim
Yeong C
YC
Ekker
Stephen C
SC
Wu
Xiaolin
X
Wang
San Ming
SM
Zhou
Xianzheng
X
D002478:Cells, Cultured; D019295:Computational Biology; D004251:DNA Transposable Elements / Q000235:genetics*; D018014:Gene Transfer Techniques; D006801:Humans; D016133:Polymerase Chain Reaction; D013601:T-Lymphocytes / Q000378:metabolism*; D019895:Transposases / Q000235:genetics / Q000378:metabolism
D004251:DNA Transposable Elements; D019895:Transposases