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Highly efficient Cas9-mediated transcriptional programming.

ABSTRACT: The RNA-guided nuclease Cas9 can be reengineered as a programmable transcription factor. However, modest levels of gene activation have limited potential applications. We describe an improved transcriptional regulator obtained through the rational design of a tripartite activator, VP64-p65-Rta (VPR), fused to nuclease-null Cas9. We demonstrate its utility in activating endogenous coding and noncoding genes, targeting several genes simultaneously and stimulating neuronal differentiation of human induced pluripotent stem cells (iPSCs).

SUBMITTER: Chavez A

PROVIDER: S-EPMC4393883 | biostudies-literature | 2015 Apr

REPOSITORIES: biostudies-literature

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Highly efficient Cas9-mediated transcriptional programming.

Chavez Alejandro A Scheiman Jonathan J Vora Suhani S Pruitt Benjamin W BW Tuttle Marcelle M P R Iyer Eswar E Lin Shuailiang S Kiani Samira S Guzman Christopher D CD Wiegand Daniel J DJ Ter-Ovanesyan Dmitry D Braff Jonathan L JL Davidsohn Noah N Housden Benjamin E BE Perrimon Norbert N Weiss Ron R Aach John J Collins James J JJ Church George M GM

Nature methods 20150302 4

The RNA-guided nuclease Cas9 can be reengineered as a programmable transcription factor. However, modest levels of gene activation have limited potential applications. We describe an improved transcriptional regulator obtained through the rational design of a tripartite activator, VP64-p65-Rta (VPR), fused to nuclease-null Cas9. We demonstrate its utility in activating endogenous coding and noncoding genes, targeting several genes simultaneously and stimulating neuronal differentiation of human ...[more]

PMID: 25730490

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