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- W2949632022 abstract "RNA plays important and diverse roles in biology,yet molecular tools to measure and manipulate RNA are limited.Recently, the bacterial adaptive immune system, CRISPR, hasrevolutionized our ability to manipulate DNA, but no knownRNA-targeting versions exist. To discover parallel bacterialRNA-targeting systems that could be used for transcriptomeengineering, we developed a computational pipeline to mine fornovel Class 2 CRISPR systems across more than 25,000 bacterialgenomes. Among the many novel CRISPR systems, we found aprogrammable RNA-targeting CRISPR system, CRISPR-Cas 13, that couldprovide immunity to E. coli against the ssRNA MS2 phage andbiochemically characterized the enzyme. We adapted CRISPR-Casl3 formodulating the transcriptome in mammalian and plant cells byheterologously expressing Casl 3 and engineering the enzyme toprecisely knockdown, bind, and edit RNA. Cas 13 knockdown was asefficient as RNA interference, but much more specific, across manytranscripts tested. RNA editing with Cas 13 was also highlyefficient, with up to 90% base editing rates, and as low as 20off-targets with engineered specificity versions. Lastly, wecombined Cas13 with isothermal amplification to develop aCRISPR-based diagnostic (CRISPR-Dx), providing rapid DNA or RNAdetection with single-molecule sensitivity and singlebase mismatchspecificity. We used this Casl3a-based molecular detectionplatform, termed SHERLOCK (Specific High Sensitivity EnzymaticReporter UnLOCKing), to specifically detect pathogenic bacteria,genotype human DNA, and identify cell-free tumor DNA mutations. Ourresults establish CRISPR-Cas13 as a flexible platform for RNAtargeting with wide applications in RNA biology, diagnostics, andtherapeutics." @default.
- W2949632022 created "2019-06-27" @default.
- W2949632022 creator A5033170303 @default.
- W2949632022 date "2018-01-01" @default.
- W2949632022 modified "2023-09-27" @default.
- W2949632022 title "Discovery of novel CRISPR enzymes for transcriptomeengineering and human health; Discovery of novel clustered regularly interspacedshort palindromic repeat enzymes for transcriptome engineering andhuman health" @default.
- W2949632022 hasPublicationYear "2018" @default.
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