CRISPR-induced DNA reorganization for multiplexed nucleic acid detection.
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
17 03 2023
17 03 2023
Historique:
received:
28
01
2022
accepted:
17
02
2023
entrez:
18
3
2023
pubmed:
19
3
2023
medline:
22
3
2023
Statut:
epublish
Résumé
Nucleic acid sensing powered by the sequence recognition of CRIPSR technologies has enabled major advancement toward rapid, accurate and deployable diagnostics. While exciting, there are still many challenges facing their practical implementation, such as the widespread need for a PAM sequence in the targeted nucleic acid, labile RNA inputs, and limited multiplexing. Here we report FACT (Functionalized Amplification CRISPR Tracing), a CRISPR-based nucleic acid barcoding technology compatible with Cas12a and Cas13a, enabling diagnostic outputs based on cis- and trans-cleavage from any sequence. Furthermore, we link the activation of CRISPR-Cas12a to the expression of proteins through a Reprogrammable PAIRing system (RePAIR). We then combine FACT and RePAIR to create FACTOR (FACT on RePAIR), a CRISPR-based diagnostic, that we use to detect infectious disease in an agricultural use case: honey bee viral infection. With high specificity and accuracy, we demonstrate the potential of FACTOR to be applied to the sensing of any nucleic acid of interest.
Identifiants
pubmed: 36932065
doi: 10.1038/s41467-023-36874-6
pii: 10.1038/s41467-023-36874-6
pmc: PMC10022571
doi:
Substances chimiques
DNA
9007-49-2
Nucleic Acids
0
RNA
63231-63-0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1505Informations de copyright
© 2023. The Author(s).
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