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Highly-efficient Cas9-mediated transcriptional programming
bioRxiv (Cold Spring Harbor Laboratory)2014
Alejandro Chavez, Jonathan Scheiman, Suhani Vora, Benjamin W. Pruitt, Marcelle Tuttle, Eswar Prasad R. Iyer, Samira Kiani, Christopher D. Guzman, Daniel J. Wiegand, Dimtry Ter-Ovanesyan, Jonathan L. Braff, Noah Davidsohn, Ron Weiss, John Aach, James J. Collins, George M. Church
Abstract
ABSTRACT The RNA-guided bacterial nuclease Cas9 can be reengineered as a programmable transcription factor by a series of changes to the Cas9 protein in addition to the fusion of a transcriptional activation domain (AD) 1–5 . However, the modest levels of gene activation achieved by current Cas9 activators have limited their potential applications. Here we describe the development of an improved transcriptional regulator through the rational design of a tripartite activator, VP64-p65-Rta (VPR), fused to Cas9. We demonstrate its utility in activating expression of endogenous coding and non-coding genes, targeting several genes simultaneously and stimulating neuronal differentiation of induced pluripotent stem cells (iPSCs).
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