GTP with a thieno[3,4-d]pyrimidine core (Thieno-GTP) is a fluorescent nucleotide that is readily accepted by T7 polymerase as a faithful GTP surrogate. This highly isomorphic nucleoside triphosphate is able to initiate transcription and incorporate during elongation. Transcripts containing a 5' pppthG are successfully dephosphorylated with alkaline phosphatase and subsequently phosphorylated with T4 polynucleotide kinase, illustrating that other terminus-modifying enzymes tolerate this modification as well.
Importantly, emissive messenger RNAs, substituted with Thieno-G are recognized by the ribosome and are capable of fluorescently reporting discrete steps in real time during translation. Additionally, Thieno-GTP can be utilized to visualize mRNA localization and assess mRNA delivery independent of translation.
|Purity||≥95% by AX-HPLC|
|Extinction Coefficient||4,500 Lmol-1cm-1 at 316 nm|
|Molecular Formula||C11H16N3O14P3S (free acid)|
|Molecular Weight||539.20 g/mole (free acid)|
|Recommended Storage||-20°C or below|
|Other Name(s)||Thieno-GTP, thGTP, 2-Aminothieno[3,4-d]pyrimidine-ribonucleoside-5'-Triphosphate|
|Application||Aptamers, Epigenetics/DNA Damage, In vitro Transcription, Mutagenesis, Photocrosslinking Studies|
|Nucleotide Category||Base Modified RNA|
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Sinkeldam, Renatus W.; McCoy, Lisa S.; Shin, Dongwon; Tor, Yitzhak . Enzymatic interconversion of isomorphic fluorescent nucleosides: adenosine deaminase transforms an adenosine analogue into an inosine analogue.