Sleeping Beauty Transposition From Nonintegrating Lentivirus

被引：59

作者：

Vink, Conrad A. ^{[1
]}

Gaspar, H. Bobby ^{[1
]}

Gabriel, Richard ^{[2
,3
]}

Schmidt, Manfred ^{[2
,3
]}

McIvor, R. Scott ^{[4
]}

Thrasher, Adrian J. ^{[1
]}

Qasim, Waseem ^{[1
]}

机构：

[1] UCL, Inst Child Hlth, Mol Immunol Unit, London WC1N 1EH, England

[2] Natl Ctr Tumor Dis, Dept Translat Oncol, Heidelberg, Germany

[3] German Canc Res Ctr, Heidelberg, Germany

[4] Univ Minnesota, Inst Human Genet, Gene Therapy Program, Minneapolis, MN 55455 USA

来源：

MOLECULAR THERAPY | 2009年 / 17卷 / 07期

关键词：

STABLE GENE-TRANSFER; EXPRESSION IN-VIVO; HUMAN GENOME; HUMAN-CELLS; TRANSPOSABLE ELEMENTS; VECTOR INTEGRATION; THERAPY; MUTAGENESIS; MOUSE; DNA;

D O I：

10.1038/mt.2009.94

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

Lentiviral vectors enter cells with high efficiency and deliver stable transduction through integration into host chromosomes, but their preference for integration within actively transcribing genes means that insertional mutagenesis following disruption of host proto-oncogenes is a recognized concern. We have addressed this problem by combining the efficient cell and nuclear entry properties of HIV-1-derived lentiviral vectors with the integration profile benefits of Sleeping Beauty (SB) transposase. Importantly, this integration enzyme does not exhibit a preference for integration within active genes. We generated integrase-deficient lentiviral vectors (IDLVs) to carry SB transposon and transposase expression cassettes. IDLVs were able to deliver transient transposase expression to target cells, and episomal lentiviral DNA was found to be a suitable substrate for integration via the SB pathway. The hybrid vector system allows genomic integration of a minimal promoter-transgene cassette flanked by short SB inverted repeats (IRs) but devoid of HIV-1 long terminal repeats (LTRs) or other virus-derived sequences. Importantly, integration site analysis revealed redirection toward a profile mimicking SB-plasmid integration and away from integration within transcriptionally active genes favored by integrase-proficient lentiviral vectors (ILVs).

引用

页码：1197 / 1204

页数：8

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