基于通路特异文库的人类细胞中化学遗传学 CRISPR-Cas9 筛选
Chemical-genetic CRISPR-Cas9 screens in human cells using a pathway-specific library.
机构信息
Department of Biochemistry and Biophysics, Helen Diller Family Comprehensive Cancer Center, University of California San Francisco, San Francisco, CA 94158, USA.
Present address: Department of Cell Biology, Blavatnik Institute of Harvard Medical School, Boston, MA 02115, USA.
出版信息
STAR Protoc. 2021 Jul 28;2(3):100685. doi: 10.1016/j.xpro.2021.100685. eCollection 2021 Sep 17.
The development of CRISPR-Cas9 screening techniques coupled with chemical inhibition of specific biological processes enables high-throughput investigation into many areas of molecular biology. We present a protocol to conduct ubiquitin proteasome system-specific chemical-genetic CRISPR-Cas9 screens in the human HAP1 cell line. This protocol can be adapted for use in other cell lines, with other compounds and types of treatments, and with any other sgRNA library. For complete details on the use and execution of this protocol, please refer to Hundley et al. (2021).
CRISPR-Cas9 筛选技术的发展加上对特定生物过程的化学抑制作用，使高通量研究分子生物学的许多领域成为可能。我们提出了一种在人类 HAP1 细胞系中进行泛素蛋白酶体系统特异性化学遗传 CRISPR-Cas9 筛选的方案。该方案可以适应于其他细胞系、其他化合物和治疗类型，以及任何其他 sgRNA 文库。有关此方案的使用和执行的完整详细信息，请参阅 Hundley 等人（2021 年）。
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