| 000 | 02203 am a22003013u 4500 | ||
|---|---|---|---|
| 042 | _adc | ||
| 100 | 1 | 0 |
_aFreund, Niklas _eauthor _92248 |
| 700 | 1 | 0 |
_aTaylor, Alexander I. _eauthor _91456 |
| 700 | 1 | 0 |
_aArangundy-Franklin, Sebastian _eauthor _92249 |
| 700 | 1 | 0 |
_aSubramanian, Nithya _eauthor _92250 |
| 700 | 1 | 0 |
_aPeak-Che _eauthor _91459 |
| 700 | 1 | 0 |
_aWhitaker, Amy M. _eauthor _92251 |
| 700 | 1 | 0 |
_aFreudenthal, Bret D. _eauthor _92252 |
| 700 | 1 | 0 |
_aAbramov, Mikhail _eauthor _92253 |
| 700 | 1 | 0 |
_aHerdewijn, Piet _eauthor _92254 |
| 700 | 1 | 0 |
_aHolliger, Philipp _eauthor _91460 |
| 245 | 0 | 0 | _aA two-residue nascent strand steric gate controls synthesis of 2'-O-methyl- and 2'-O-(2-methoxyethyl)-RNA |
| 260 | _c2022-10-13. | ||
| 500 | _a/pmc/articles/PMC7614059/ | ||
| 500 | _a/pubmed/36229679 | ||
| 520 | _aSteric exclusion is a key element of enzyme substrate specificity, including in polymerases. Such substrate specificity restricts the enzymatic synthesis of 2'-modified nucleic acids, which are of interest in nucleic acid-based drug development. Here we describe the discovery of a two-residue, nascent strand, steric control "gate" in an archaeal DNA polymerase. We show that engineering of the gate to reduce steric bulk in the context of a previously-described RNA polymerase activity unlocks the synthesis of 2'-modified RNA oligomers, specifically the efficient synthesis of both defined and random-sequence 2'-O-methyl-RNA (2'OMe-RNA) and 2'-O-(2-methoxyethyl)-RNA (MOE-RNA) oligomers up to 750 nt. This enabled the discovery of RNA endonuclease catalysts entirely composed of 2'OMe-RNA ("2'OMezymes") for the allele-specific cleavage of oncogenic KRAS (G12D) and β-catenin CTNNB1 (S33Y) mRNAs, and the elaboration of mixed 2'OMe- / MOE-RNA aptamers with high affinity for Vascular Endothelial Growth Factor. Our results open up these 2'-modified RNAs-used in several approved nucleic acid therapeutics-for enzymatic synthesis and a wider exploration in directed evolution and nanotechnology. | ||
| 540 | _a | ||
| 546 | _aen | ||
| 690 | _aArticle | ||
| 655 | 7 |
_aText _2local |
|
| 786 | 0 | _nNat Chem | |
| 856 | 4 | 1 |
_uhttp://dx.doi.org/10.1038/s41557-022-01050-8 _zConnect to this object online. |
| 999 |
_c1893 _d1893 |
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