RNA-guided gene activation by CRISPR-Cas9-based transcription factors

By P. Perez-Pinera, D. D. Koçak, Christopher M. Vockley, Andrew F Adler, A. Kabadi, Lauren R. Polstein, Pratiksha I. Thakore, Katherine A. Glass, David G. Ousterout, Kam W. Leong and 4 more. Nature Methods, 2013

1,302 citations recorded by the source corpus.

Technologies for engineering synthetic transcription factors have enabled many advances in medical and scientific research. In contrast to existing methods based on engineering of DNA-binding proteins, we created a Cas9-based transactivator that is targeted to DNA sequences by guide RNA molecules. Coexpression of this transactivator and combinations of guide RNAs in human cells induced specific expression of endogenous target genes, demonstrating a simple and versatile approach for RNA-guided gene activation.

Subfields: Molecular Biology

View Paper: https://www.semanticscholar.org/paper/369525574750ff5d7a4de74342610ebe7ca879e8

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In Molecular Biology

Citations in the corpus

Development and Applications of CRISPR-Cas9 for Genome Engineering (2014, positive): “Although it is still unclear how Cas9 is affected by chromatin accessibility and heterochromatin versus euchromatin, dCas9 transcriptional activators can upregulate transcription at sites lacking DNase I hypersensitivity sites, indicating successful binding to inaccessible chromatin (Perez-Pinera et”
Repurposing the CRISPR-Cas9 system for targeted DNA methylation (2016, background): “Finally, the choice of the targeting domain for both genome and epigenome editing seems to be shifting toward the CRISPR-Cas9 system (8,15), which is not surprising because of the ease of targeting by guide RNAs even to multiple sites (25) and insensitivity to CpG methylation.”
CHOPCHOP v3: expanding the CRISPR web toolbox beyond genome editing (2019, methodology): “For instance, CRISPR–Cas has been used to introduce new sequences into the genome (1–3), activate (4,5) or repress (6) transcription, as an enrichment tool for sequencing (7,8), for targeted hypermutation (9), as a diagnostic tool (10), to perform whole-organism lineage tracing (11), to target RNA m”
A protein tagging system for signal amplification in gene expression and fluorescence imaging (2014, positive): “…copies of dCas9-VP64 to a native promoter using multiple nonoverlap- 642 Cell 159, 635–646, October 23, 2014 ª2014 Elsevier Inc. ping sgRNAs also greatly enhanced transcriptional activation (Cheng et al., 2013; Hu et al., 2014; Maeder et al., 2013; Mali et al., 2013; Perez-Pinera et al., 2013).”