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Gene Transfer Efficiency and Genome-Wide Integration Profiling of Sleeping Beauty, Tol2, and PiggyBac Transposons in Human Primary T Cells

  1. Author:
    Huang, X.
    Guo, H. F.
    Tammana, S.
    Jung, Y. C.
    Mellgren, E.
    Bassi, P.
    Cao, Q.
    Tu, Z. J.
    Kim, Y. C.
    Ekker, S. C.
    Wu, X. L.
    Wang, S. M.
    Zhou, X. Z.

  2. Author Address

    [Huang, Xin; Guo, Hongfeng; Zhou, Xianzheng] Univ Minnesota, Sch Med, Masonic Canc Ctr, Minneapolis, MN 55455 USA. [Huang, Xin; Guo, Hongfeng; Zhou, Xianzheng] Univ Minnesota, Sch Med, Div Pediat Blood & Marrow Transplantat, Minneapolis, MN 55455 USA. [Huang, Xin; Guo, Hongfeng; Zhou, Xianzheng] Univ Minnesota, Sch Med, Ctr Immunol, Minneapolis, MN 55455 USA. [Huang, Xin; Guo, Hongfeng; Zhou, Xianzheng] Univ Minnesota, Sch Med, Ctr Genome Engn, Minneapolis, MN 55455 USA. [Tammana, Syam; Zhou, Xianzheng] Univ Minnesota, Grad Sch, Grad Program Microbial Engn, Minneapolis, MN 55455 USA. [Jung, Yong-Chul; Kim, Yeong C.; Wang, San Ming] Northwestern Univ, Evanston NW Healthcare Res Inst, Ctr Funct Genom, Evanston, IL USA. [Mellgren, Emil] Macalester Coll, St Paul, MN 55105 USA. [Bassi, Preetinder] Univ Minnesota, Coll Biol Sci, Minneapolis, MN 55455 USA. [Cao, Qing] Univ Minnesota, Sch Med, Biostat & Informat Core Masonic Canc Ctr, Minneapolis, MN 55455 USA. [Tu, Zheng Jin] Univ Minnesota, Inst Supercomp, Minneapolis, MN 55455 USA. [Ekker, Stephen C.] Mayo Clin, Rochester, MN USA. [Wu, Xiaolin] Natl Canc Inst Frederick, Lab Mol Technol, SAIC Frederick Inc, Frederick, MD USA.;Zhou, XZ, Univ Minnesota, Sch Med, Masonic Canc Ctr, MMC 366,420 Delaware St, Minneapolis, MN 55455 USA.;zhoux058@umn.edu
    1. Year: 2010
    2. Date: Oct
  2. Journal: Molecular Therapy
    1. 18
    2. 10
    3. Pages: 1803-1813
  4. Type of Article: Article
  5. ISSN: 1525-0016
  1. 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 DNasel 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.

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External Sources

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  2. DOI: 10.1038/mt.2010.141
  3. View record in Web of Science®
  4. WOS: 000282541200011

Library Notes

  1. Fiscal Year: FY2010-2011
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