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Debranchase-resistant labeling of RNA using the 10DM24 deoxyribozyme and fluorescent modified nucleotides.

ABSTRACT: The 10DM24 deoxyribozyme can site-specifically label RNAs with fluorophore-GTP conjugates; however, the 2',5'-branched RNA linkage is readily cleaved by debranchase. To prevent loss of labels upon cleavage, we synthesized phosphorothioate-modified, fluorescent GTP derivatives and elaborated conditions for their incorporation by 10DM24. RNAs labeled with fluorescent derivatives of Sp-GTPS were found to be resistant to debranchase.

SUBMITTER: Carrocci TJ

PROVIDER: S-EPMC5668183 | biostudies-literature | 2017 Nov

REPOSITORIES: biostudies-literature

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Debranchase-resistant labeling of RNA using the 10DM24 deoxyribozyme and fluorescent modified nucleotides.

Carrocci Tucker J TJ Lohe Lea L Ashton Matthew J MJ Höbartner Claudia C Hoskins Aaron A AA

Chemical communications (Cambridge, England) 20171006 88

The 10DM24 deoxyribozyme can site-specifically label RNAs with fluorophore-GTP conjugates; however, the 2',5'-branched RNA linkage is readily cleaved by debranchase. To prevent loss of labels upon cleavage, we synthesized phosphorothioate-modified, fluorescent GTP derivatives and elaborated conditions for their incorporation by 10DM24. RNAs labeled with fluorescent derivatives of Sp-GTP<sup>S</sup> were found to be resistant to debranchase. ...[more]

PMID: 28984884

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