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Amino-Functionalized 5′ Cap Analogs as Tools for Site-Specific Sequence-Independent Labeling of mRNA

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cris.lastimport.scopus2024-02-12T20:20:08Z
dc.abstract.enmRNA is a template for protein biosynthesis, and consequently mRNA transport, translation, and turnover are key elements in the overall regulation of gene expression. Along with growing interest in the mechanisms regulating mRNA decay and localization, there is an increasing need for tools enabling convenient fluorescent labeling or affinity tagging of mRNA. We report new mRNA 5′ cap analog-based tools that enable site-specific labeling of RNA within the cap using N-hydroxysuccinimide (NHS) chemistry. We explored two complementary methods: a co-transcriptional labeling method, in which the label is first attached to a cap analog and then incorporated into RNA by in vitro transcription, and a post-transcriptional labeling method, in which an amino-functionalized cap analog is incorporated into RNA followed by chemical labeling of the resulting transcript. After testing the biochemical properties of RNAs carrying the novel modified cap structures, we demonstrated the utility of fluorescently labeled RNAs in decapping assays, RNA decay assays, and RNA visualization in cells. Finally, we also demonstrated that mRNAs labeled by the reported method are translationally active. We envisage that the novel analogs will provide an alternative to radiolabeling of mRNA caps for in vitro studies and open possibilities for new applications related to the study of mRNA fates in vivo.
dc.affiliationUniwersytet Warszawski
dc.contributor.authorSikorski, Paweł
dc.contributor.authorWarmińska, Zofia
dc.contributor.authorKropiwnicka, Anna
dc.contributor.authorŁukaszewicz, Maciej
dc.contributor.authorDarżynkiewicz, Edward
dc.contributor.authorŻuberek, Joanna
dc.contributor.authorJemielity, Jacek
dc.contributor.authorKowalska, Joanna
dc.contributor.authorWarmiński, Marcin
dc.date.accessioned2024-01-24T16:28:42Z
dc.date.available2024-01-24T16:28:42Z
dc.date.issued2017
dc.description.financeNie dotyczy
dc.identifier.doi10.1021/ACS.BIOCONJCHEM.7B00291
dc.identifier.issn1043-1802
dc.identifier.urihttps://repozytorium.uw.edu.pl//handle/item/100543
dc.languageeng
dc.pbn.affiliationbiological sciences
dc.relation.ispartofBioconjugate Chemistry
dc.rightsClosedAccess
dc.sciencecloudnosend
dc.titleAmino-Functionalized 5′ Cap Analogs as Tools for Site-Specific Sequence-Independent Labeling of mRNA
dc.typeJournalArticle
dspace.entity.typePublication