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Enzymatic synthesis of hypermodified DNA polymers for sequence-specific display of four different hydrophobic groups
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SYSNO ASEP 0537186 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Enzymatic synthesis of hypermodified DNA polymers for sequence-specific display of four different hydrophobic groups Author(s) Ondruš, Marek (UOCHB-X) ORCID, RID
Sýkorová, Veronika (UOCHB-X) ORCID, RID
Bednárová, Lucie (UOCHB-X) RID, ORCID
Pohl, Radek (UOCHB-X) RID, ORCID
Hocek, Michal (UOCHB-X) RID, ORCIDSource Title Nucleic Acids Research. - : Oxford University Press - ISSN 0305-1048
Roč. 48, č. 21 (2020), s. 11982-11993Number of pages 12 s. Language eng - English Country GB - United Kingdom Keywords nucleotides ; DNA ; functionalized polymers Subject RIV CE - Biochemistry OECD category Biochemistry and molecular biology R&D Projects GX20-00885X GA ČR - Czech Science Foundation (CSF) Method of publishing Open access Institutional support UOCHB-X - RVO:61388963 UT WOS 000606018700018 EID SCOPUS 85098456968 DOI 10.1093/nar/gkaa999 Annotation A set of modified 2'-deoxyribonucleoside triphosphates (dNTPs) bearing a linear or branched alkane, indole or phenyl group linked through ethynyl or alkyl spacer were synthesized and used as substrates for polymerase synthesis of hypermodified DNA by primer extension (PEX). Using the alkyl-linked dNTPs, the polymerase synthesized up to 22-mer fully modified oligonucleotide (ON), whereas using the ethynyl-linked dNTPs, the enzyme was able to synthesize even long sequences of >100 modified nucleotides in a row. In PCR, the combinations of all four modified dNTPs showed only linear amplification. Asymmetric PCR or PEX with separation or digestion of the template strand can be used for synthesis of hypermodified single-stranded ONs, which are monodispersed polymers displaying four different substituents on DNA backbone in sequence-specific manner. The fully modified ONs hybridized with complementary strands and modified DNA duplexes were found to exist in B-type conformation (B- or C-DNA) according to CD spectral analysis. The modified DNA can be replicated with high fidelity to natural DNA through PCR and sequenced. Therefore, this approach has a promising potential in generation and selection of hypermodified aptamers and other functional polymers. 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 2021 Electronic address https://doi.org/10.1093/nar/gkaa999
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