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Zhou_X

Research Abstracts Online
January 2009 - March 2010

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University of Minnesota Twin Cities
Academic Health Center
Masonic Cancer Center

PI: Xianzheng Zhou

Transposon Integration Profiling in Human Hematopeietic Cells

DNA transposons have emerged as a useful tool in gene therapy. However, their integration site preferences remain elusive. This study directly compares the genomic integration efficiencies and transposition site preferences of Sleeping Beauty (SB), Tol2, and piggyBac (PB) in primary T cells derived from peripheral blood (PBL) and umbilical cord blood (UCB). PB demonstrated the highest efficiency stable gene transfer in PBL-derived T cells, whereas SB and Tol2 mediated intermediate and lowest efficiencies, respectively. Southern blot analysis of ~132 T-cell clones demonstrated that more identical clones were found in PB and Tol2 transfected T cells derived from PBL and UCB but not in SB transfected T cells. T-cell receptor Vb analyses showed that PB and Tol2 may drive clonal expansion in CD4 and CD8 T-cell subsets. Genome-wide integration analysis demonstrated that SB, Tol2 and PB integrations occurred in all the chromosomes with no preference. Additionally, Tol2 and PB integration sites were mainly localized near the transcriptional start sites, CpG islands and DNase hypersensitive sites, whereas SB integrations were randomly distributed. These results suggest that SB might be safer than Tol2 and PB in T-cell engineering and support continuing development of an SB-mediated T-cell Phase I trial.

Group Members

Xin Huang, Research Associate
Changming Lu, Research Associate