title: Gene transfer efficiency and genome-wide integration profiling of Sleeping Beauty, Tol2, and piggyBac transposons in human primary T cells.
abstract: 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.
authors.affiliation: Division of Pediatric Blood and Marrow Transplantation, University of Minnesota Medical School, Minneapolis, Minnesota, USA.
authors.forename: Xin
authors.initials: X
authors.lastname: Huang
authors.forename: Hongfeng
authors.initials: H
authors.lastname: Guo
authors.forename: Syam
authors.initials: S
authors.lastname: Tammana
authors.forename: Yong-Chul
authors.initials: YC
authors.lastname: Jung
authors.forename: Emil
authors.initials: E
authors.lastname: Mellgren
authors.forename: Preetinder
authors.initials: P
authors.lastname: Bassi
authors.forename: Qing
authors.initials: Q
authors.lastname: Cao
authors.forename: Zheng Jin
authors.initials: ZJ
authors.lastname: Tu
authors.forename: Yeong C
authors.initials: YC
authors.lastname: Kim
authors.forename: Stephen C
authors.initials: SC
authors.lastname: Ekker
authors.forename: Xiaolin
authors.initials: X
authors.lastname: Wu
authors.forename: San Ming
authors.initials: SM
authors.lastname: Wang
authors.forename: Xianzheng
authors.initials: X
authors.lastname: Zhou
mesh_terms: 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
chemical_list: D004251:DNA Transposable Elements; D019895:Transposases