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Reprogrammed CRISPR-Cas13b suppresses SARS-CoV-2 replication and circumvents its mutational escape through mismatch tolerance
- Source :
- Nature Communications, Nature Communications, Vol 12, Iss 1, Pp 1-16 (2021)
- Publication Year :
- 2021
- Publisher :
- Nature Publishing Group UK, 2021.
-
Abstract
- The recent dramatic appearance of variants of concern of SARS-coronavirus-2 (SARS-CoV-2) highlights the need for innovative approaches that simultaneously suppress viral replication and circumvent viral escape from host immunity and antiviral therapeutics. Here, we employ genome-wide computational prediction and single-nucleotide resolution screening to reprogram CRISPR-Cas13b against SARS-CoV-2 genomic and subgenomic RNAs. Reprogrammed Cas13b effectors targeting accessible regions of Spike and Nucleocapsid transcripts achieved >98% silencing efficiency in virus-free models. Further, optimized and multiplexed Cas13b CRISPR RNAs (crRNAs) suppress viral replication in mammalian cells infected with replication-competent SARS-CoV-2, including the recently emerging dominant variant of concern B.1.1.7. The comprehensive mutagenesis of guide-target interaction demonstrated that single-nucleotide mismatches does not impair the capacity of a potent single crRNA to simultaneously suppress ancestral and mutated SARS-CoV-2 strains in infected mammalian cells, including the Spike D614G mutant. The specificity, efficiency and rapid deployment properties of reprogrammed Cas13b described here provide a molecular blueprint for antiviral drug development to suppress and prevent a wide range of SARS-CoV-2 mutants, and is readily adaptable to other emerging pathogenic viruses.<br />Cas13b can be harnessed to target and degrade RNA transcripts inside a cellular environment. Here the authors reprogram Cas13b to target SARSCoV-2 transcripts in infected mammalian cells and reveal its resilience to variants thanks to single mismatch tolerance.
- Subjects :
- RNAi therapy
CRISPR-Cas systems
medicine.drug_class
Science
viruses
General Physics and Astronomy
Mutagenesis (molecular biology technique)
Computational biology
Genome, Viral
Biology
Virus Replication
Antiviral Agents
General Biochemistry, Genetics and Molecular Biology
Article
03 medical and health sciences
0302 clinical medicine
Drug Development
RNA interference
Chlorocebus aethiops
medicine
Gene silencing
CRISPR
Animals
Humans
Clustered Regularly Interspaced Short Palindromic Repeats
Vero Cells
Synthetic biology
030304 developmental biology
Subgenomic mRNA
Trans-activating crRNA
0303 health sciences
Multidisciplinary
SARS-CoV-2
fungi
COVID-19
General Chemistry
COVID-19 Drug Treatment
HEK293 Cells
Viral replication
RNAi
Mutation
Spike Glycoprotein, Coronavirus
Antiviral drug
030217 neurology & neurosurgery
Subjects
Details
- Language :
- English
- ISSN :
- 20411723
- Volume :
- 12
- Database :
- OpenAIRE
- Journal :
- Nature Communications
- Accession number :
- edsair.doi.dedup.....c06af7239b0ba8fb210223d3c97693e3