A drug-inducible transgenic system for direct reprogramming of multiple somatic cell types.
A drug-inducible transgenic system for direct reprogramming of multiple somatic cell types.
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DOI：
10.1038/nbt1483
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发表时间：
2008-08
影响因子：
46.9
通讯作者：
Jaenisch, Rudolf
中科院分区：
文献类型：
--
作者：
Wernig, Marius;Lengner, Christopher J.;Hanna, Jacob;Lodato, Michael A.;Steine, Eveline;Foreman, Ruth;Staerk, Judith;Markoulaki, Styliani;Jaenisch, Rudolf
诱导多能性的研究由于需要高滴度逆转录病毒载体的感染而变得复杂，这导致遗传异质性的细胞群。我们产生了遗传上同质的“次生”体细胞，这些体细胞携带重编程因子，被定义为强力霉素（dox）诱导的转基因。这些细胞是通过用dox诱导的慢病毒感染成纤维细胞，加入dox重新编程，选择诱导多能干细胞和产生嵌合小鼠而产生的。来自这些嵌合体的细胞在还原暴露后无需病毒感染即可重编程，其效率比使用直接感染和药物选择进行多能性标记再激活观察到的效率高25- 50倍。我们证明(i)不同诱导水平的重编程因子可以诱导多能性，（ii）转基因活性的持续时间与重编程效率直接相关，（iii）来自许多体细胞组织的细胞可以被重编程，（iv）不同类型的细胞需要不同的诱导水平。该系统有助于表征重编程，并为基因或化学筛选提供工具，以增强重编程。
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.
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