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EPR Distance Measurements on Long Non-coding RNAs Empowered by Genetic Alphabet Expansion Transcription.

ABSTRACT: We present herein a novel nitroxide spin label-containing RNA triphosphate TPT3NO and its application for site-specific spin-labeling of RNA through in?vitro transcription using an expanded genetic alphabet. Our strategy allows the facile preparation of spin-labeled RNAs with sizes ranging from short RNA oligonucleotides to large, complex RNA molecules with over 370 nucleotides by standard in?vitro transcription. As a proof of concept, inter-spin distance distributions are measured by pulsed electron paramagnetic resonance (EPR) spectroscopy in short self-complementary RNA sequences and in a well-studied 185?nucleotide non-coding RNA, the B.?subtilis glmS ribozyme. The approach is then applied to probe for the first time the folding of the 377?nucleotide A-region of the long non-coding RNA Xist, by PELDOR.

SUBMITTER: Domnick C

PROVIDER: S-EPMC7318606 | biostudies-literature | 2020 May

REPOSITORIES: biostudies-literature

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EPR Distance Measurements on Long Non-coding RNAs Empowered by Genetic Alphabet Expansion Transcription.

Domnick Christof C Eggert Frank F Wuebben Christine C Bornewasser Lisa L Hagelueken Gregor G Schiemann Olav O Kath-Schorr Stephanie S

Angewandte Chemie (International ed. in English) 20200313 20

We present herein a novel nitroxide spin label-containing RNA triphosphate TPT3<sup>NO</sup> and its application for site-specific spin-labeling of RNA through in vitro transcription using an expanded genetic alphabet. Our strategy allows the facile preparation of spin-labeled RNAs with sizes ranging from short RNA oligonucleotides to large, complex RNA molecules with over 370 nucleotides by standard in vitro transcription. As a proof of concept, inter-spin distance distributions are measured by ...[more]

PMID: 31981397

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