%0 Journal Article %1 Jiaoeabe7106 %A Jiao, Chunlei %A Sharma, Sahil %A Dugar, Gaurav %A Peeck, Natalia L. %A Bischler, Thorsten %A Wimmer, Franziska %A Yu, Yanying %A Barquist, Lars %A Schoen, Christoph %A Kurzai, Oliver %A Sharma, Cynthia M. %A Beisel, Chase L. %D 2021 %I American Association for the Advancement of Science %J Science %K myown %N 6545 %P 941-948 %R 10.1126/science.abe7106 %T Noncanonical crRNAs derived from host transcripts enable multiplexable RNA detection by Cas9 %U https://science.sciencemag.org/content/early/2021/04/26/science.abe7106 %V 372 %X 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 SARS-CoV-2 and its D614G variant with single-base resolution in patient samples.

@article{Jiaoeabe7106, abstract = {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 SARS-CoV-2 and its D614G variant with single-base resolution in patient samples.}, added-at = {2021-04-27T17:00:39.000+0200}, author = {Jiao, Chunlei and Sharma, Sahil and Dugar, Gaurav and Peeck, Natalia L. and Bischler, Thorsten and Wimmer, Franziska and Yu, Yanying and Barquist, Lars and Schoen, Christoph and Kurzai, Oliver and Sharma, Cynthia M. and Beisel, Chase L.}, biburl = {https://www.bibsonomy.org/bibtex/2288be302dd3f313986329fc633ff6331/cusysmed}, day = 27, description = {Noncanonical crRNAs derived from host transcripts enable multiplexable RNA detection by Cas9 | Science}, doi = {10.1126/science.abe7106}, elocation-id = {eabe7106}, eprint = {https://science.sciencemag.org/content/early/2021/04/26/science.abe7106.full.pdf}, interhash = {58cb4179207c5c1cefe4e96077a8ca5a}, intrahash = {288be302dd3f313986329fc633ff6331}, issn = {0036-8075}, journal = {Science}, keywords = {myown}, month = apr, number = 6545, pages = {941-948}, publisher = {American Association for the Advancement of Science}, timestamp = {2024-04-16T17:15:17.000+0200}, title = {Noncanonical crRNAs derived from host transcripts enable multiplexable RNA detection by Cas9}, url = {https://science.sciencemag.org/content/early/2021/04/26/science.abe7106}, volume = 372, year = 2021 }