The expanded development and application of CRISPR system for sensitive nucleotide detection
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Protein & Cell
COMMENTARY The expanded development and application of CRISPR system for sensitive nucleotide detection Fengjing Jia1, Xuewen Li4, Chao Zhang1&, Xueming Tang2,3& Translational Medical Center for Stem Cell Therapy and Institute for Regenerative Medicine, Shanghai East Hospital, Shanghai Key Laboratory of Signaling and Disease Research, School of Life Sciences and Technology, Tongji University, Shanghai 200000, China 2 Institute of Biotechnology Research, Shanghai Academy of Agricultural Sciences, Key Laboratory of Agricultural Genetics and Breeding, Shanghai 201106, China 3 Crops Ecological Environment Security Inspection and Supervision Center (Shanghai), Ministry of Agriculture and Rural Affairs, Shanghai 201106, China 4 Silicon Gene Tech Co., Ltd., Shanghai 200124, China & Correspondence: [email protected] (C. Zhang), [email protected] (X. Tang)
CRISPR/Cas system, originally developed as genetic editing tool, also shows great potentials for nucleotide detection. A recent study published in Molecular Cell (Freije et al., 2019) developed a Cas13a-based CARVER (Cas13-assisted restriction of viral expression and readout) to detect RNA viruses such as lymphocytic choriomeningitis, influenza A and vesicular stomatitis, which provided a potential expanded application for the detection of a broad range of viral nucleotides in disease diagnosis. CRISPR/Cas (clustered regularly interspaced short palindromic repeats/CRISPR-associated) systems are utilized by bacteria and archaea as adaptive immune system to defend against phage infection. Cas effectors are guided by a CRISPR RNAs (crRNAs) to bind and cut DNA or RNA targets to defend against invading nucleotides (Horvath and Barrangou, 2010; Sorek et al., 2013; Barrangou and Marraffini, 2014). The discovery of CRISPR/Cas system dated back to 1987, the regularly spaced direct repeats were first found in the iap gene of Escherichia coli (Ishino et al., 1987). Until 2002, the spaced direct repeats were named as CRISPR (Jansen et al., 2002). In 2012, Jinek et al. reported that CRISPR/Cas9 could specifically cleave the target DNA with a single RNA chimera (Jinek et al., 2012), which opened the prelude of CRISPR/Cas9 system for genomic editing. Since CRISPR/Cas9 was discovered, CRISPR/Cas systems attracted much attention and CRISPR toolbox had been continuously expanded. As a potent complement to DNA targeting CRISPR toolbox, CRISPR/Cas12a (previously known as CpfI), a Class 2 type V CRISPR/Cas effector, was characterized (Zetsche et al., 2015) with the
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capability to efficiently cleave target double-stranded DNA (dsDNA) guided by a crRNA. Moreover, differing from Cas9, Cas12a possessed a target-dependent nonspecific singlestranded DNA (ssDNA) cutting activity (Chen et al., 2018). Beyond dsDNA, ssRNA molecules could also be edited by another Cas protein, CRISPR/Cas13a (previously known as C2c2) (Abudayyeh et al., 2016). Cas13a, as a class 2 type VI CRISPR effector, was programmed to cleave the target RNA guided by crRNA. In addition, as
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