引导RNA
RNA编辑
基因
非编码RNA
RNA剪接
亚基因组mRNA
信使核糖核酸
基因组
抄写(语言学)
遗传学
作者
Chunlei Jiao,Sahil Sharma,Gaurav Dugar,Natalia L. Peeck,Thorsten Bischler,Franziska Wimmer,Yanying Yu,Lars Barquist,Christoph Schoen,Oliver Kurzai,Cynthia M. Sharma,Chase L. Beisel
出处
期刊:Science
[American Association for the Advancement of Science (AAAS)]
日期:2021-05-28
卷期号:372 (6545): 941-948
被引量:25
标识
DOI:10.1126/science.abe7106
摘要
CRISPR-Cas systems recognize foreign genetic material using CRISPR RNAs (crRNAs). In type II systems, a trans-activating crRNA (tracrRNA) hybridizes to crRNAs to drive their processing and utilization by Cas9. While analyzing Cas9-RNA complexes from Campylobacter jejuni, we discovered tracrRNA hybridizing to cellular RNAs, leading to formation of noncanonical crRNAs capable of guiding DNA targeting by Cas9. Our discovery inspired the engineering of reprogrammed tracrRNAs that link the presence of any RNA of interest to DNA targeting with different Cas9 orthologs. This capability became the basis for a multiplexable diagnostic platform termed LEOPARD (leveraging engineered tracrRNAs and on-target DNAs for parallel RNA detection). LEOPARD allowed simultaneous detection of RNAs from different viruses in one test and distinguished severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and its D614G (Asp614→Gly) variant with single-base resolution in patient samples.
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