期刊
ACS CHEMICAL BIOLOGY
卷 13, 期 2, 页码 406-416出版社
AMER CHEMICAL SOC
DOI: 10.1021/acschembio.7b00657
关键词
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资金
- NIH/NIGMS [DP2 GM119139]
- NIH/NINDS [U54 NS100717]
- NIH/NCI [K99/R00 CA181494]
- Allen Distinguished Investigator Award
- Paul F. Glenn Center for Aging Research, a Calico/QB3 Longevity Award
- Tau Consortium
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 elucidating 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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