Targeted Epigenetic Remodeling of Endogenous Loci by CRISPR/Cas9-Based Transcriptional Activators Directly Converts Fibroblasts to Neuronal Cells.
Targeted Epigenetic Remodeling of Endogenous Loci by CRISPR/Cas9-Based Transcriptional Activators Directly Converts Fibroblasts to Neuronal Cells.
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DOI：
10.1016/j.stem.2016.07.001
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发表时间：
2016-09-01
期刊：
影响因子：
23.9
通讯作者：
Gersbach, Charles A.
中科院分区：
文献类型：
--
作者：
Black, Joshua B.;Adler, Andrew F.;Wang, Hong-Gang;D'Ippolito, Anthony M.;Hutchinson, Hunter A.;Reddy, Timothy E.;Pitt, Geoffrey S.;Leong, Kam W.;Gersbach, Charles A.
外源命运决定转录因子的过度表达可以直接将分化的体细胞重新编程为靶细胞类型。在这里，我们表明，类似的重新编程也可以通过使用基于CRISPR/Cas9的工程转录激活剂直接激活内源基因来实现。我们使用这种方法来诱导内源性Brn2、Ascl1和Myt1基因(BAM因子)的激活，以将小鼠胚胎成纤维细胞转化为诱导的神经细胞。这种内源基因的直接激活迅速重塑了目标基因座的表观遗传状态，并在重新编程过程中诱导了内源基因的持续表达。因此，内源性主要转录因子的转录激活和表观遗传重塑足以实现细胞类型之间的转换。通过这种方法在其天然染色质环境中快速和持续地激活内源基因，可能有助于使用避免基因组整合的瞬时方法进行重新编程，并为克服表观遗传障碍对细胞命运的指定提供了一种新的策略。布莱克等人。研究表明，通过基于CRISPR/Cas9的内源性神经源性基因的激活，成纤维细胞重新编程为诱导的神经元，导致目标内源性基因位点的快速表观遗传重塑，并在重新编程过程中保持基因表达。
Overexpression of exogenous fate-specifying transcription factors can directly reprogram differentiated somatic cells to target cell types. Here, we show that similar reprogramming can also be achieved through the direct activation of endogenous genes using engineered CRISPR/Cas9-based transcriptional activators. We use this approach to induce activation of the endogenous Brn2, Ascl1, and Myt1l genes (BAM factors) to convert mouse embryonic fibroblasts to induced neuronal cells. This direct activation of endogenous genes rapidly remodeled the epigenetic state of the target loci and induced sustained endogenous gene expression during reprogramming. Thus, transcriptional activation and epigenetic remodeling of endogenous master transcription factors is sufficient for conversion between cell types. The rapid and sustained activation of endogenous genes in their native chromatin context by this approach may facilitate reprogramming with transient methods that avoid genomic integration and provides a new strategy for overcoming epigenetic barriers to cell fate specification. Black et al. show that reprogramming of fibroblasts to induced neurons via CRISPR/Cas9-based activation of endogenous neurogenic genes leads to rapid epigenetic remodeling at the targeted endogenous loci and sustained gene expression throughout the reprogramming process.
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影响因子：
46.9
作者：
通讯作者：
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影响因子：
64.8
作者：
通讯作者：
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DOI：
10.1038/mtna.2012.25
发表时间：
2012-07-10
期刊：
Molecular therapy. Nucleic acids
影响因子：
--
作者：
通讯作者：
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影响因子：
56.9
作者：
Jinek, Martin;Chylinski, Krzysztof;Charpentier, Emmanuelle
通讯作者：
Charpentier, Emmanuelle
DOI：
10.1126/science.1232033
发表时间：
2013-02-15
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Mali P;Yang L;Esvelt KM;Aach J;Guell M;DiCarlo JE;Norville JE;Church GM
通讯作者：
Church GM