Number of the records: 1
Expedient production of site specifically nucleobase-labelled or hypermodified RNA with engineered thermophilic DNA polymerases
- 1.
SYSNO ASEP 0585094 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Expedient production of site specifically nucleobase-labelled or hypermodified RNA with engineered thermophilic DNA polymerases Author(s) Brunderová, Mária (UOCHB-X) ORCID
Havlíček, Vojtěch (UOCHB-X)
Matyašovský, Ján (UOCHB-X) ORCID, RID
Pohl, Radek (UOCHB-X) RID, ORCID
Poštová Slavětínská, Lenka (UOCHB-X) RID
Krömer, Matouš (UOCHB-X) RID, ORCID
Hocek, Michal (UOCHB-X) RID, ORCIDArticle number 3054 Source Title Nature Communications. - : Nature Publishing Group
Roč. 15, č. 1 (2024)Number of pages 13 s. Language eng - English Country US - United States R&D Projects GX20-00885X GA ČR - Czech Science Foundation (CSF) EH22_008/0004575 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) Method of publishing Open access Institutional support UOCHB-X - RVO:61388963 UT WOS 001202403000013 EID SCOPUS 85189966748 DOI 10.1038/s41467-024-47444-9 Annotation Innovative approaches to controlled nucleobase-modified RNA synthesis are urgently needed to support RNA biology exploration and to synthesize potential RNA therapeutics. Here we present a strategy for enzymatic construction of nucleobase-modified RNA based on primer-dependent engineered thermophilic DNA polymerases – SFM4-3 and TGK. We demonstrate introduction of one or several different base-modified nucleotides in one strand including hypermodified RNA containing all four modified nucleotides bearing four different substituents, as well as strategy for primer segment removal. We also show facile site-specific or segmented introduction of fluorophores or other functional groups at defined positions in variety of RNA molecules, including structured or long mRNA. Intriguing translation efficacy of single-site modified mRNAs underscores the necessity to study isolated modifications placed at designer positions to disentangle their biological effects and enable development of improved mRNA therapeutics. Our toolbox paves the way for more precise dissecting RNA structures and functions, as well as for construction of diverse types of base-functionalized RNA for therapeutic applications and diagnostics. Workplace Institute of Organic Chemistry and Biochemistry Contact asep@uochb.cas.cz ; Kateřina Šperková, Tel.: 232 002 584 ; Jana Procházková, Tel.: 220 183 418 Year of Publishing 2025 Electronic address https://doi.org/10.1038/s41467-024-47444-9
Number of the records: 1