使用基于CRISPR-Cpf1的转录因子进行可诱导和多重基因调控。
Inducible and multiplex gene regulation using CRISPR-Cpf1-based transcription factors.
作者信息
Tak Y Esther, Kleinstiver Benjamin P, Nuñez James K, Hsu Jonathan Y, Horng Joy E, Gong Jingyi, Weissman Jonathan S, Joung J Keith
机构信息
Molecular Pathology Unit, Center for Cancer Research, and Center for Computational and Integrative Biology, Massachusetts General Hospital, Charlestown, Massachusetts, USA.
Department of Pathology, Harvard Medical School, Boston, Massachusetts, USA.
出版信息
Nat Methods. 2017 Dec;14(12):1163-1166. doi: 10.1038/nmeth.4483. Epub 2017 Oct 30.
Targeted and inducible regulation of mammalian gene expression is a broadly important capability. We engineered drug-inducible catalytically inactive Cpf1 nuclease fused to transcriptional activation domains to tune the expression of endogenous genes in human cells. Leveraging the multiplex capability of the Cpf1 platform, we demonstrate both synergistic and combinatorial gene expression in human cells. Our work should enable the development of multiplex gene perturbation library screens for understanding complex cellular phenotypes.
对哺乳动物基因表达进行靶向和诱导调控是一项具有广泛重要性的能力。我们设计了与转录激活结构域融合的药物诱导型催化失活Cpf1核酸酶，以调节人类细胞中内源基因的表达。利用Cpf1平台的多重功能，我们在人类细胞中展示了协同和组合基因表达。我们的工作应能推动多重基因扰动文库筛选的发展，以了解复杂的细胞表型。
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