Protected 5-(hydroxymethyl) uracil nucleotides bearing visible-light photocleavable groups as building blocks for polymerase synthesis of photocaged DNA

0489970 - ÚOCHB 2019 RIV GB eng J - Journal Article
Boháčová, Soňa - Ludvíková, L. - Poštová Slavětínská, Lenka - Vaníková, Zuzana - Klán, P. - Hocek, Michal
Protected 5-(hydroxymethyl) uracil nucleotides bearing visible-light photocleavable groups as building blocks for polymerase synthesis of photocaged DNA.
Organic & Biomolecular Chemistry. Roč. 16, č. 9 (2018), s. 1527-1535. ISSN 1477-0520. E-ISSN 1477-0539
R&D Projects: GA ČR(CZ) GA17-03419S
Grant - others:AV ČR(CZ) AP1501
Program: Akademická prémie - Praemium Academiae
Institutional support: RVO:61388963
Keywords : stem cell DNA * restriction endonucleases * mammalian cells
OECD category: Organic chemistry
Impact factor: 3.490, year: 2018
http://pubs.rsc.org/en/content/articlehtml/2018/ob/c8ob00160j

Nucleosides, nucleotides and 2'-deoxyribonucleoside triphosphates (dNTPs) containing 5-(hydroxy-methyl) uracil protected with photocleavable groups (2-nitrobenzyl-, 6-nitropiperonyl or 9-anthrylmethyl) were prepared and tested as building blocks for the polymerase synthesis of photocaged oligonucleotides and DNA. Photodeprotection (photorelease) reactions were studied in detail on model nucleoside mono-phosphates and their photoreaction quantum yields were determined. Photocaged dNTPs were then tested and used as substrates for DNA polymerases in primer extension or PCR. DNA probes containing photocaged or free 5-hydroxymethylU in the recognition sequence of restriction endonucleases were prepared and used for the study of photorelease of caged DNA by UV or visible light at different wavelengths. The nitropiperonyl-protected nucleotide was found to be a superior building block because the corresponding dNTP is a good substrate for DNA polymerases, and the protecting group is efficiently cleavable by irradiation by UV or visible light (up to 425 nm).
Permanent Link: http://hdl.handle.net/11104/0284265