An iCRISPR platform for rapid, multiplexable, and inducible genome editing in human pluripotent stem cells.
An iCRISPR platform for rapid, multiplexable, and inducible genome editing in human pluripotent stem cells.
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一个 iCRISPR 平台，用于在人类多能干细胞中进行快速、可多重、可诱导的基因组编辑。
DOI：
10.1016/j.stem.2014.05.018
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发表时间：
2014-08-07
期刊：
影响因子：
23.9
通讯作者：
Huangfu, Danwei
中科院分区：
文献类型：
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作者：
Gonzalez, Federico;Zhu, Zengrong;Shi, Zhong-Dong;Lelli, Katherine;Verma, Nipun;Li, Qing V.;Huangfu, Danwei
人类多能干细胞（hPSCs）为阐明复杂性状和疾病背后的基因和分子途径提供了一个独特的平台。为了实现这一希望，迫切需要快速可控的基因操作方法。通过结合两种新开发的基因编辑工具，TALEN和CRISPR/Cas系统，我们开发了一种新的hPSCs基因组工程平台，我们将其命名为iCRISPR。iCRISPR能够快速高效地生成用于功能丧失研究的双等位基因敲除hPSCs，以及具有特定核苷酸改变的纯合子敲除hPSCs，用于精确建模疾病状况。我们进一步证明，首次有效的一步产生双基因和三基因敲除hPSC系，以及阶段特异性诱导基因敲除在hPSC分化。因此，iCRISPR平台特别适合于人类疾病研究中复杂遗传相互作用和多效性基因功能的解剖，并具有支持高通量hPSCs遗传分析的潜力。
Human pluripotent stem cells (hPSCs) offer a unique platform for elucidating the genes and molecular pathways that underlie complex traits and diseases. To realize this promise, methods for rapid and controllable genetic manipulations are urgently needed. By combining two newly developed gene-editing tools, the TALEN and CRISPR/Cas systems, we have developed a novel genome-engineering platform in hPSCs, which we named iCRISPR. iCRISPR enabled rapid and highly efficient generation of biallelic knockout hPSCs for loss-of-function studies, as well as homozygous knockin hPSCs with specific nucleotide alterations for precise modeling of disease conditions. We further demonstrate, for the first time, efficient one-step generation of double- and triple-gene knockout hPSC lines, as well as stage-specific inducible gene knockout during hPSC differentiation. Thus the iCRISPR platform is uniquely suited for dissection of complex genetic interactions and pleiotropic gene functions in human disease studies, and has the potential to support high-throughput genetic analysis in hPSCs.
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影响因子：
7
作者：
Cho SW;Kim S;Kim Y;Kweon J;Kim HS;Bae S;Kim JS
通讯作者：
Kim JS
影响因子：
46.9
作者：
Hockemeyer, Dirk;Soldner, Frank;Beard, Caroline;Gao, Qing;Mitalipova, Maisam;DeKelver, Russell C.;Katibah, George E.;Amora, Ranier;Boydston, Elizabeth A.;Zeitler, Bryan;Meng, Xiangdong;Miller, Jeffrey C.;Zhang, Lei;Rebar, Edward J.;Gregory, Philip D.;Urnov, Fyodor D.;Jaenisch, Rudolf
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Jaenisch, Rudolf
影响因子：
56.9
作者：
Jinek, Martin;Chylinski, Krzysztof;Charpentier, Emmanuelle
通讯作者：
Charpentier, Emmanuelle
影响因子：
46.9
作者：
Cho, Seung Woo;Kim, Sojung;Kim, Jin-Soo
通讯作者：
Kim, Jin-Soo
影响因子：
46.9
作者：
通讯作者：
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