Most cited article - PubMed ID 39748090
Enzymatic synthesis of reactive RNA probes containing squaramate-linked cytidine or adenosine for bioconjugations and cross-linking with lysine-containing peptides and proteins
A series of adenosine triphosphate (ATP) derivatives bearing chloro, fluoro, amino, methyl, vinyl, and ethynyl groups at position 2 are synthesized and tested as substrates for RNA and DNA polymerases. The modified nucleotides work well in in vitro transcription with T7 RNA polymerase and primer extension (PEX) using engineered DNA polymerases (TGK, 2M) except for the bulkier 2-vinyl- and 2-ethynyl-ATP derivatives that give truncated products. However, in single nucleotide incorporation followed by PEX, they still can be used for site-specific incorporation of reactive modifications into RNA that can be further used for postsynthetic labeling through thiol-ene or Cu-catalyzed alkyne-azide cycloadditions reactions. All modified ATPs work in polyadenylation catalyzed by poly(A) polymerase to form long 3'-polyA tails containing the modifications that also can be used for labeling.
- Keywords
- DNA polymerases, RNA polymerases, click reactions, nucleosides triphosphates, nucleotides, polyA polymerase, thiol‐ene addition,
- MeSH
- Adenosine Triphosphate * chemical synthesis chemistry analogs & derivatives metabolism MeSH
- Cycloaddition Reaction MeSH
- DNA-Directed RNA Polymerases * metabolism chemistry MeSH
- DNA-Directed DNA Polymerase * metabolism chemistry MeSH
- RNA * chemistry metabolism MeSH
- Viral Proteins metabolism MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- Adenosine Triphosphate * MeSH
- bacteriophage T7 RNA polymerase MeSH Browser
- DNA-Directed RNA Polymerases * MeSH
- DNA-Directed DNA Polymerase * MeSH
- RNA * MeSH
- Viral Proteins MeSH
