TY  - BOOK
AU  - Taylor,Alexander I.
AU  - Wan,Christopher J.K.
AU  - Donde,Maria J.
AU  - Peak-Che
AU  - Holliger,Philipp
TI  - A modular XNAzyme cleaves long, structured RNAs under physiological conditions and enables allele-specific gene silencing
PY  - 2022///-11
KW  - Text
KW  - local
N1  - /pmc/articles/PMC7613789; /pubmed/36064973
N2  - Nucleic acid catalysts (ribozymes, DNA- and XNAzymes) cleave target (m)RNAs with high specificity but have shown limited efficacy in clinical application. 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-β-D-arabino nucleic acid (FANA). FR6_1 overcomes 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 Mg(2+) 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 knockdown 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
UR  - http://dx.doi.org/10.1038/s41557-022-01021-z
ER  -