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Pruitt, Benjamin W.

Publications and source records attributed to Pruitt, Benjamin W..

4 records
All sourcesNASA NTRSDOE OSTI
DOE OSTI2015

Cas9 gRNA engineering for genome editing, activation and repression

Here we demonstrate that by altering the length of Cas9-associated guide RNA(gRNA) we were able to control Cas9 nuclease activity and simultaneously perform genome editing and transcriptional regulation with a single Cas9 protein. We exploited these principles to engineer mammalian synthetic circuits with combined transcriptional regulation and kill functions governed by a single multifunctional Cas9 protein.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2015

Highly efficient Cas9-mediated transcriptional programming

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

59 BASIC BIOLOGICAL SCIENCES↗