Highly efficient Cas9-mediated transcriptional programming
Open Access
- 2 March 2015
- journal article
- research article
- Published by Springer Science and Business Media LLC in Nature Methods
- Vol. 12 (4), 326-328
- https://doi.org/10.1038/nmeth.3312
Abstract
The fusion of three transcriptional activation domains to a nuclease-deficient Cas9 achieves robust induction of gene expression and can induce differentiation of hiPSCs. 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).Keywords
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