CRISPR RNA-guided activation of endogenous human genes

By Morgan L. Maeder, S. J. Linder, Vincent M Cascio, Yanfang Fu, Quan H Ho, J. Keith Joung. Nature Methods, 2013

1,215 citations recorded by the source corpus.

Short guide RNAs (gRNAs) can direct catalytically inactive CRISPR-associated 9 nuclease (dCas9) to repress endogenous genes in bacteria and human cells. Here we show that single or multiple gRNAs can direct dCas9 fused to a VP64 transcriptional activation domain to increase expression of endogenous human genes. This proof-of-principle work shows that clustered regularly interspaced short palindromic repeat (CRISPR)-Cas systems can target heterologous effector domains to endogenous sites in human cells.

Subfields: Molecular Biology

View Paper: https://www.semanticscholar.org/paper/308047767e5092abfa954fccdbe0cf2c4e740e54

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Citations in the corpus

Genome-wide binding of the CRISPR endonuclease Cas9 in mammalian cells (2014, neutral): “The catalytically inactive dCas9 (D10A and H840A mutations) alone or when fused to activators or repressors has been used to modulate transcription 20–25 and dCas9 has also been fused to GFP to allow imaging of genomic loci in living cells 26 .”
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”
Compact and highly active next-generation libraries for CRISPR-mediated gene repression and activation (2016, background): “, 2013) to programmably interfere with (CRISPRi) or activate (CRISPRa) transcription (Gilbert et al., 2014, 2013; Konermann et al., 2015; Maeder et al., 2013; PerezPinera et al., 2013; Tanenbaum et al., 2014), and used these to systematically manipulate gene”
A CRISPR/Cas9 Toolbox for Multiplexed Plant Genome Editing and Transcriptional Regulation1[OPEN] (2015, background): “…such as VP16 or VP64 (Sadowski et al., 1988; Beerli et al., 1998) to a deactivated Cas9 (dCas9) lacking endonuclease activity can up-regulate endogenous gene expression in human cells (Cheng et al., 2013; Gilbert et al., 2013; Maeder et al., 2013; Mali et al., 2013a; Perez-Pinera et al., 2013).”
Dimeric CRISPR RNA-guided FokI nucleases for highly specific genome editing (2014, methodology): “Our Csy4-based system should also prove generally useful for any application requiring the expression of two or more gRNAs (e.g.—the targeting of multiple dCas9-activator proteins to achieve synergistic upregulation of gene expression 20–22 ).”
Crystal Structure of Cas9 in Complex with Guide RNA and Target DNA (2014, background): “Moreover, a catalytically inactive Cas9 can serve as an RNA-guided DNA-binding platform to target effector domains and modulate endogenous transcription (Gilbert et al., 2013; Konermann et al., 2013; Maeder et al., 2013; Perez-Pinera et al., 2013; Qi et al., 2013).”