000			02104 am a22002533u 4500
042			_adc
100	1	0	_aTaylor, Alexander I. _eauthor _91456
700	1	0	_aWan, Christopher J.K. _eauthor _91457
700	1	0	_aDonde, Maria J. _eauthor _91458
700	1	0	_aPeak-Che _eauthor _91459
700	1	0	_aHolliger, Philipp _eauthor _91460
245	0	0	_aA modular XNAzyme cleaves long, structured RNAs under physiological conditions and enables allele-specific gene silencing
260			_c2022-11.
500			_a/pmc/articles/PMC7613789/
500			_a/pubmed/36064973
520			_aNucleic 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.
540			_a
540			_ahttps://www.springernature.com/gp/open-research/policies/accepted-manuscript-termsUsers may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use: https://www.springernature.com/gp/open-research/policies/accepted-manuscript-terms
546			_aen
690			_aArticle
655	7		_aText _2local
786	0		_nNat Chem
856	4	1	_uhttp://dx.doi.org/10.1038/s41557-022-01021-z _zConnect to this object online.
999			_c1952 _d1952