Highly-efficient Cas9-mediated transcriptional programming
By Alejandro Chavez, J. Scheiman, Suhani Vora, Benjamin W Pruitt, Marcelle Tuttle, Eswar P. R. Iyer, Shuailiang Lin, S. Kiani, Christopher D Guzman, Daniel J. Wiegand and 9 more. 2014
1,473 citations recorded by the source corpus.
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).
Subfields: Aging, Molecular Biology
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In Molecular Biology
- Lei S. Qi
- Haifeng Wang
- Marie La Russa
- Luke A. Gilbert
- Jonathan S. Weissman
- H. Rees
- Max A. Horlbeck
- Britt Adamson
Citations in the corpus
- Evolved Cas9 variants with broad PAM compatibility and high DNA specificity (2018, positive): “Endogenous gene activation was done similarly but with 200 ng of dCas9–VPR plasmid and 50 ng of sgRNA expression plasmid only.”
- Highly Specific Epigenome Editing by CRISPR/Cas9 Repressors for Silencing of Distal Regulatory Elements (2015, positive): “In the CRISPR/dCas9 system adapted from Streptococcus pyogenes, co-delivery of a single guide RNA (sgRNA) directs dCas9 binding through an 18–20 nucleotide protospacer region complementary to the target genomic sequence 13–15 .”
- Compact and highly active next-generation libraries for CRISPR-mediated gene repression and activation (2016, background): “Importantly, while several strategies have been described for CRISPR-mediated activation (Chavez et al., 2015; Gilbert et al., 2013; Hilton et al., 2015; Konermann et al., 2015; Maeder et al., 2013; PerezPinera et al., 2013; Tanenbaum et al., 2014; Zalatan et al., 2015), a recent comparison of…”
- CRISPR/Cas9 in Genome Editing and Beyond. (2016, methodology): “All of these studies show that synergistically recruiting multiple activators to the dCas9 target locus enhances the activation of the CRISPRa system (180, 184, 185).”