Science (American Association for the Advancement of Science), 2021-05, Vol.372 (6545), p.941-948
2021
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Details
- Autor(en) / Beteiligte
- Titel
- Noncanonical crRNAs derived from host transcripts enable multiplexable RNA detection by Cas9
- Ist Teil von
- Science (American Association for the Advancement of Science), 2021-05, Vol.372 (6545), p.941-948
- Ort / Verlag
- United States: The American Association for the Advancement of Science
- Erscheinungsjahr
- 2021
- Quelle
- American Association for the Advancement of Science
- Beschreibungen/Notizen
- 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 , 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 (Asp →Gly) variant with single-base resolution in patient samples.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 0036-8075eISSN: 1095-9203DOI: 10.1126/science.abe7106Titel-ID: cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_8224270
- Format
- –
- Schlagworte
- Base Sequence, Biomarkers, Campylobacter jejuni, Clustered Regularly Interspaced Short Palindromic Repeats, Coronaviruses, COVID-19 - diagnosis, COVID-19 - virology, COVID-19 Nucleic Acid Testing, CRISPR, CRISPR-Associated Protein 9 - metabolism, CRISPR-Cas Systems, Deoxyribonucleic acid, DNA, Genome editing, Genomes, Humans, Matching, Microbio, Molec Biol, Multiplexing, Nuclease, Nucleic Acid Hybridization, Respiratory diseases, Ribonucleic acid, RNA, RNA - analysis, RNA - genetics, RNA, Bacterial - genetics, RNA, Guide, CRISPR-Cas Systems - genetics, RNA, Messenger - genetics, RNA, Viral - analysis, RNA, Viral - genetics, SARS-CoV-2 - genetics, Severe acute respiratory syndrome, Severe acute respiratory syndrome coronavirus 2, Spike Glycoprotein, Coronavirus - genetics, Viral diseases, Viruses