Cas9 as a versatile tool for engineering biology

By P. Mali, Kevin M. Esvelt, George M. Church. Nature Methods, 2013

1,228 citations recorded by the source corpus.

RNA-guided Cas9 nucleases derived from clustered regularly interspaced short palindromic repeats (CRISPR)-Cas systems have dramatically transformed our ability to edit the genomes of diverse organisms. We believe tools and techniques based on Cas9, a single unifying factor capable of colocalizing RNA, DNA and protein, will grant unprecedented control over cellular organization, regulation and behavior. Here we describe the Cas9 targeting methodology, detail current and prospective engineering advances and suggest potential applications ranging from basic science to the clinic.

Subfields: Molecular Biology

View Paper: https://www.semanticscholar.org/paper/b2f075f3cb9715bc42237fb5a730de88d45b9ec9

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Citations in the corpus

Genome-wide binding of the CRISPR endonuclease Cas9 in mammalian cells (2014, positive): “The simplicity of targeting any locus with a single protein and a programmable sgRNA has quickly led to widespread use of Cas9 11,12 in applications such as genome editing 7,8,13–16 , disease gene repair 17,18 and knock-in of specific tags 8,19 .”
Point-of-care CRISPR/Cas nucleic acid detection: Recent advances, challenges and opportunities (2020, neutral): “This is only the case for the protospacer present on the viral DNA and not for the spacer on the CRISPR array (Mali et al., 2013).”
Discovery and functional characterization of diverse Class 2 CRISPR-Cas systems (2015, background): “These systems have recently attracted much attention due to their unique, “Lamarckian” mode of action that retains “memories” from past infections and provides specific resistance to these infections via an RNA-guided process that has been harnessed to create powerful genome editing tools (Cong et a”
Concerning RNA-guided gene drives for the alteration of wild populations (2014, background): “engineered, or evolved Cas9 variants with improved properties (Esvelt et al., 2011; Mali et al., 2013b).”
Mechanisms and Regulation of Alternative Pre-mRNA Splicing (2015, background): “A variety of therapeutic strategies, such as small molecules (40, 41) and antisense oligonucleotides (42–44), as well as genome editing using CRISPR/Cas9 (45, 46), show promise for future intervention to ameliorate the diseasing-causing effects of human mutations on patterns of alternative splicing.”
CRISPR/Cas9-mediated gene editing in human tripronuclear zygotes (2015, background): “…containing the tri-nucleotide protospacer adjacent motif (PAM) and complementary to the gRNA, and can be programmed to recognize virtually any genes through the manipulation of gRNA sequences (Cho et al., 2013; Cong et al., 2013; Jinek et al., 2012; Jinek et al., 2013; Mali et al., 2013c).”