Vigilant: An Engineered VirD2-Cas9 Complex for Lateral Flow Assay-Based Detection of SARS-CoV2.

COVID-19 CRISPR-Cas9 RT-RPA SARS-CoV-2 VirD2 biosensors dCas9 lateral flow assay molecular diagnostics nucleic acid detection relaxases

Journal

Nano letters
ISSN: 1530-6992
Titre abrégé: Nano Lett
Pays: United States
ID NLM: 101088070

Informations de publication

Date de publication:
28 04 2021
Historique:
pubmed: 13 4 2021
medline: 25 6 2021
entrez: 12 4 2021
Statut: ppublish

Résumé

Rapid, sensitive, and specific point-of-care testing for pathogens is crucial for disease control. Lateral flow assays (LFAs) have been employed for nucleic acid detection, but they have limited sensitivity and specificity. Here, we used a fusion of catalytically inactive SpCas9 endonuclease and VirD2 relaxase for sensitive, specific nucleic acid detection by LFA. In this assay, the target nucleic acid is amplified with biotinylated oligos. VirD2-dCas9 specifically binds the target sequence via dCas9 and covalently binds to a FAM-tagged oligonucleotide via VirD2. The biotin label and FAM tag are detected by a commercially available LFA. We coupled this system, named Vigilant (

Identifiants

pubmed: 33844549
doi: 10.1021/acs.nanolett.1c00612
pmc: PMC8056947
doi:

Substances chimiques

RNA, Viral 0

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

3596-3603

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Auteurs

Tin Marsic (T)

Laboratory for Genome Engineering and Synthetic Biology, Division of Biological Sciences, King Abdullah University of Science and Technology (KAUST), Thuwal 23955-6900, Saudi Arabia.

Zahir Ali (Z)

Laboratory for Genome Engineering and Synthetic Biology, Division of Biological Sciences, King Abdullah University of Science and Technology (KAUST), Thuwal 23955-6900, Saudi Arabia.

Muhammad Tehseen (M)

Laboratory of DNA Replication and Recombination, Biological and Environmental Sciences and Engineering Division, King Abdullah University of Science and Technology (KAUST), Thuwal 23955-6900, Saudi Arabia.

Ahmed Mahas (A)

Laboratory for Genome Engineering and Synthetic Biology, Division of Biological Sciences, King Abdullah University of Science and Technology (KAUST), Thuwal 23955-6900, Saudi Arabia.

Samir Hamdan (S)

Laboratory of DNA Replication and Recombination, Biological and Environmental Sciences and Engineering Division, King Abdullah University of Science and Technology (KAUST), Thuwal 23955-6900, Saudi Arabia.

Magdy Mahfouz (M)

Laboratory for Genome Engineering and Synthetic Biology, Division of Biological Sciences, King Abdullah University of Science and Technology (KAUST), Thuwal 23955-6900, Saudi Arabia.

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Classifications MeSH