Noncanonical crRNAs derived from host transcripts enable multiplexable RNA detection by Cas9.
Science
; 372(6545): 941-948, 2021 05 28.
Article
in English
| MEDLINE | ID: covidwho-1205995
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 severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and its D614G (Asp614âGly) variant with single-base resolution in patient samples.
Full text:
Available
Collection:
International databases
Database:
MEDLINE
Main subject:
RNA
/
RNA, Viral
/
RNA, Guide, Kinetoplastida
/
Clustered Regularly Interspaced Short Palindromic Repeats
/
CRISPR-Associated Protein 9
/
SARS-CoV-2
Type of study:
Diagnostic study
Topics:
Variants
Limits:
Humans
Language:
English
Journal:
Science
Year:
2021
Document Type:
Article
Affiliation country:
Science.abe7106
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