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Inducible, tunable and multiplex human gene regulation using CRISPR-Cpf1-based transcription factors

By Yu Gyoung Tak, Benjamin P. Kleinstiver, James K. Nuñez, Jonathan Y Hsu, Jingyi Gong, Jonathan S. Weissman, J. Keith Joung

Posted 15 Jun 2017
bioRxiv DOI: 10.1101/150656

Targeted and inducible regulation of mammalian gene expression is a broadly important research capability that may also enable development of novel therapeutics for treating human diseases. Here we demonstrate that a catalytically inactive RNA-guided CRISPR-Cpf1 nuclease fused to transcriptional activation domains can up-regulate endogenous human gene expression. We engineered drug-inducible Cpf1-based activators and show how this system can be used to tune the regulation of endogenous gene transcription in human cells. Leveraging the simpler multiplex capability of the Cpf1 platform, we show that we can induce both synergistic and combinatorial gene expression in human cells. Our work should enable the creation of other Cpf1-based gene regulatory fusion proteins and the development of multiplex gene perturbation library screens for understanding complex cellular phenotypes.

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