期刊
NATURE CHEMISTRY
卷 14, 期 11, 页码 1295-+出版社
NATURE PORTFOLIO
DOI: 10.1038/s41557-022-01021-z
关键词
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资金
- Medical Research Council [MC_U105178804]
- Biotechnology and Biological Sciences Research Council [INTENSIFY BB/M005623/1]
- AMGEN Scholars programme
- Wellcome Trust
- Wellcome Trust/Royal Society Sir Henry Dale Fellowship [215453/Z/19/Z]
- Wellcome Trust [215453/Z/19/Z] Funding Source: Wellcome Trust
This study reports the development of a highly specific RNA endonuclease XNAzyme, FR6_1, that can cleave highly structured full-length mRNA with allelic selectivity for tumor-associated mutations. This research provides a starting point for the development of improved gene-silencing agents based on FANA or other XNA chemistries.
Nucleic-acid catalysts (ribozymes, DNA- and XNAzymes) cleave target (m)RNAs with high specificity but have shown limited efficacy in clinical applications. Here we report on the in vitro evolution and engineering of a highly specific modular RNA endonuclease XNAzyme, FR6_1, composed of 2'-deoxy-2'-fluoro-beta-D-arabino nucleic acid (FANA). FR6_1 overcomes the activity limitations of previous DNA- and XNAzymes and can be retargeted to cleave highly structured full-length (>5 kb) BRAF and KRAS mRNAs at physiological Mg2+ concentrations with allelic selectivity for tumour-associated (BRAF V600E and KRAS G12D) mutations. Phosphorothioate-FANA modification enhances FR6_1 biostability and enables rapid KRAS mRNA knock-down in cultured human adenocarcinoma cells with a G12D-allele-specific component provided by in vivo XNAzyme cleavage activity. These results provide a starting point for the development of improved gene-silencing agents based on FANA or other XNA chemistries.
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