CRISPRi and CRISPRa Screens in Mammalian Cells for Precision Biology and Medicine,ACS Chemical Biology

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CRISPRi and CRISPRa Screens in Mammalian Cells for Precision Biology and Medicine
ACS Chemical Biology ( IF 3.5 ) Pub Date : 2017-10-24 00:00:00 , DOI: 10.1021/acschembio.7b00657
Martin Kampmann _{1,

2}

Affiliation

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.

中文翻译：

用于精确生物学和医学的哺乳动物细胞中的CRISPRi和CRISPRa筛选

下一代DNA测序技术已导致来自患者和健康个体的基因组和转录组数据的大量积累。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意义。哺乳动物细胞中的遗传筛选是系统地阐明健康和疾病状态下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。这篇综述的重点是将CRISPR干扰（CRISPRi）和CRISPR激活（CRISPRa）用于哺乳动物细胞的遗传筛选。我们介绍了基础技术，并介绍了不同类型的CRISPRi / a筛选器，包括基于细胞存活/增殖，对药物或毒素的敏感性，荧光报告基因和单细胞转录组的那些。以大量基因对为靶标以构建遗传相互作用图的组合筛选，揭示了途径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核酸酶的筛选技术。最后，我们强调未来的挑战和机遇。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核酸酶的筛选技术。最后，我们强调了未来的挑战和机遇。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核酸酶的筛选技术。最后，我们强调未来的挑战和机遇。

更新日期：2017-10-25

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