Programmable modulation of ribosomal frameshifting by mRNA targeting CRISPR-Cas12a system.

Autor:	Huang SH; Department of Biochemistry and Molecular Biology, College of Medicine, National Cheng Kung University, Tainan, Taiwan.; Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, Tainan, Taiwan., Chen SC; Department of Biochemistry and Molecular Biology, College of Medicine, National Cheng Kung University, Tainan, Taiwan.; National Institute of Cancer Research, National Health Research Institutes, Tainan, Taiwan., Wu TY; YD BioLabs, Inc., Hsinchu, Taiwan., Chen CY; YD BioLabs, Inc., Hsinchu, Taiwan.; School of Medical Laboratory Science and Biotechnology, Taipei Medical University, Taipei, Taiwan., Yu CH; Department of Biochemistry and Molecular Biology, College of Medicine, National Cheng Kung University, Tainan, Taiwan.; Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, Tainan, Taiwan.
Jazyk:	angličtina
Zdroj:	IScience [iScience] 2023 Nov 19; Vol. 26 (12), pp. 108492. Date of Electronic Publication: 2023 Nov 19 (Print Publication: 2023).
DOI:	10.1016/j.isci.2023.108492
Abstrakt:	Minus 1 programmed ribosomal frameshifting (-1 PRF) is a conserved translational regulation event essential for critical biological processes, including the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) replication. Efficient trans -modulation of the structured RNA element crucial to -1 PRF will endow the therapeutic application. Here, we demonstrate that CRISPR RNA can stimulate efficient -1 PRF. Assembled CRISPR-Cas12a, but not CRISPR-Cas9, complex further enhances -1 PRF efficiency through its higher capacity to stall translating ribosomes. We additionally perform CRISPR-Cas12a targeting to impair the SARS-CoV-2 frameshifting pseudoknot structure via a focused screening. We demonstrate that targeting CRISPR-Cas12a results in more than 70% suppression of -1 PRF in vitro and about 50% suppression in mammalian cells. Our results show the expanded function of the CRISPR-Cas12 system in modulating -1 PRF efficiency through stalling ribosomes and deforming frameshifting stimulatory signals, which could serve as a new strategy for future coronavirus pandemics. Competing Interests: C.-Y.C. is the co-founder and shareholder of YD Biolabs, Ltd. T.-Y.W. is an employee of YD Biolabs, Ltd. (© 2023 The Author(s).)
Databáze:	MEDLINE
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