| Article ID | Journal | Published Year | Pages | File Type |
|---|---|---|---|---|
| 2093674 | Stem Cell Reports | 2014 | 8 Pages |
•RNA-guided VP64dCas9-BFPVP64 fusion protein robustly activates endogenous Myod1•Transactivated Myod1 can reprogram mouse embryonic fibroblasts to skeletal myocytes•VP64 fusion to both the N and C terminus of dCas9-BFP facilitates reprogramming•Myogenic gene expression is comparable to MYOD1 overexpression-based reprogramming
SummaryGene activation by the CRISPR/Cas9 system has the potential to enable new approaches to science and medicine, but the technology must be enhanced to robustly control cell behavior. We show that the fusion of two transactivation domains to Cas9 dramatically enhances gene activation to a level that is necessary to reprogram cell phenotype. Targeted activation of the endogenous Myod1 gene locus with this system led to stable and sustained reprogramming of mouse embryonic fibroblasts into skeletal myocytes. The levels of myogenic marker expression obtained by the activation of endogenous Myod1 gene were comparable to that achieved by overexpression of lentivirally delivered MYOD1 transcription factor.
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