A drug-inducible transgenic system for direct reprogramming of multiple somatic cell types

被引：326

作者：

Wernig, Marius ^{[1
]}

Lengner, Christopher J. ^{[1
]}

Hanna, Jacob ^{[1
]}

Lodato, Michael A. ^{[1
,2
]}

Steine, Eveline ^{[1
]}

Foreman, Ruth ^{[1
,2
]}

Staerk, Judith ^{[1
]}

Markoulaki, Styliani ^{[1
]}

Jaenisch, Rudolf ^{[1
,2
]}

机构：

[1] Whitehead Inst Biomed Res, Cambridge, MA 02142 USA

[2] MIT, Dept Biol, Cambridge, MA 02139 USA

来源：

NATURE BIOTECHNOLOGY | 2008年 / 26卷 / 08期

关键词：

D O I：

10.1038/nbt1483

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：

The study of induced pluripotency is complicated by the need for infection with high-titer retroviral vectors, which results in genetically heterogeneous cell populations. We generated genetically homogeneous 'secondary' somatic cells that carry the reprogramming factors as defined doxycycline (dox)-inducible transgenes. These cells were produced by infecting fibroblasts with dox-inducible lentiviruses, reprogramming by dox addition, selecting induced pluripotent stem cells and producing chimeric mice. Cells derived from these chimeras reprogram upon dox exposure without the need for viral infection with efficiencies 25- to 50-fold greater than those observed using direct infection and drug selection for pluripotency marker reactivation. We demonstrate that (i) various induction levels of the reprogramming factors can induce pluripotency, (ii) the duration of transgene activity directly correlates with reprogramming efficiency, (iii) cells from many somatic tissues can be reprogrammed and (iv) different cell types require different induction levels. This system facilitates the characterization of reprogramming and provides a tool for genetic or chemical screens to enhance reprogramming.

引用

页码：916 / 924

页数：9

共 28 条

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