CRISPRi and CRISPRa Screens in Mammalian Cells for Precision Biology and Medicine.
CRISPRi and CRISPRa Screens in Mammalian Cells for Precision Biology and Medicine.
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DOI：
10.1021/acschembio.7b00657
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发表时间：
2018-02-16
影响因子：
4
通讯作者：
Kampmann M
中科院分区：
文献类型：
--
作者：
Kampmann M
下一代DNA测序技术已经从患者和健康个体中积累了大量的基因组和转录组数据。未来的主要挑战是了解人类基因组和转录组元素的功能意义，以及对诊断和治疗的影响。哺乳动物细胞的遗传筛选是系统地阐明健康和疾病状态中基因功能的有力方法。特别是，最近开发的基于CRISPR/ cas9的筛选方法在揭示人类疾病的机制和治疗策略方面具有巨大的潜力。本综述的重点是在哺乳动物细胞中使用CRISPR干扰（CRISPRi）和CRISPR激活（CRISPRa）进行遗传筛选。我们介绍了基础技术，并介绍了不同类型的CRISPRi/a筛选，包括基于细胞存活/增殖、对药物或毒素的敏感性、荧光报告和单细胞转录组的筛选。组合筛选，其中大量的基因对为目标，以构建遗传相互作用图谱，揭示通路关系和蛋白质复合物。我们将CRISPRi和CRISPRa与其他技术进行比较和对比，包括RNA干扰（RNAi）和基于CRISPR核酸酶的筛选。最后，我们强调了未来的挑战和机遇。
Next-generation DNA sequencing technologies have led to a massive accumulation of genomic and transcriptomic data from patients and healthy individuals. The major challenge ahead is to understand the functional significance of the elements of the human genome and transcriptome, and implications for diagnosis and treatment. Genetic screens in mammalian cells are a powerful approach to systematically elucidate gene function in health and disease states. In particular, recently developed CRISPR/Cas9-based screening approaches have enormous potential to uncover mechanisms and therapeutic strategies for human diseases. The focus of this review is the use of CRISPR interference (CRISPRi) and CRISPR activation (CRISPRa) for genetic screens in mammalian cells. We introduce the underlying technology and present different types of CRISPRi/a screens, including those based on cell survival/proliferation, sensitivity to drugs or toxins, fluorescent reporters, and single-cell transcriptomes. Combinatorial screens, in which large numbers of gene pairs are targeted to construct genetic interaction maps, reveal pathway relationships and protein complexes. We compare and contrast CRISPRi and CRISPRa with alternative technologies, including RNA interference (RNAi) and CRISPR nuclease-based screens. Finally, we highlight challenges and opportunities ahead.
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影响因子：
48
作者：
Chavez A;Tuttle M;Pruitt BW;Ewen-Campen B;Chari R;Ter-Ovanesyan D;Haque SJ;Cecchi RJ;Kowal EJK;Buchthal J;Housden BE;Perrimon N;Collins JJ;Church G
通讯作者：
Church G
影响因子：
50.3
作者：
Berger AH;Brooks AN;Wu X;Shrestha Y;Chouinard C;Piccioni F;Bagul M;Kamburov A;Imielinski M;Hogstrom L;Zhu C;Yang X;Pantel S;Sakai R;Watson J;Kaplan N;Campbell JD;Singh S;Root DE;Narayan R;Natoli T;Lahr DL;Tirosh I;Tamayo P;Getz G;Wong B;Doench J;Subramanian A;Golub TR;Meyerson M;Boehm JS
通讯作者：
Boehm JS
影响因子：
7.7
作者：
Acosta-Alvear D;Cho MY;Wild T;Buchholz TJ;Lerner AG;Simakova O;Hahn J;Korde N;Landgren O;Maric I;Choudhary C;Walter P;Weissman JS;Kampmann M
通讯作者：
Kampmann M
影响因子：
46.9
作者：
Evers, Bastiaan;Jastrzebski, Katarzyna;Bernards, Rene
通讯作者：
Bernards, Rene
影响因子：
46.9
作者：
Cho, Seung Woo;Kim, Sojung;Kim, Jin-Soo
通讯作者：
Kim, Jin-Soo