Chemo‐enzymatic treatment of RNA to facilitate analyses

• Published in: Wiley interdisciplinary reviews. RNA 2020-01, Vol.11 (1), p.e1561-n/a
• Main Authors: Muthmann, Nils, Hartstock, Katja, Rentmeister, Andrea
• Format: Article
• Language: English
• Subjects: Animals; chemo‐enzymatic; click chemistry; Enzymes - metabolism; Gene expression; Humans; metabolic labeling; Metabolism; Next-generation sequencing; Nucleotide sequence; RNA - analysis; RNA - genetics; RNA - metabolism; RNA editing; RNA labeling; RNA modification; RNA processing; Staining and Labeling
• ISSN: 1757-7004; 1757-7012
• DOI: 10.1002/wrna.1561

Labeling RNA is a recurring problem to make RNA compatible with state‐of‐the‐art methodology and comes in many flavors. Considering only cellular applications, the spectrum still ranges from site‐specific labeling of individual transcripts, for example, for live‐cell imaging of mRNA trafficking, to metabolic labeling in combination with next generation sequencing to capture dynamic aspects of RNA metabolism on a transcriptome‐wide scale. Combining the specificity of RNA‐modifying enzymes with non‐natural substrates has emerged as a valuable strategy to modify RNA site‐ or sequence‐specifically with functional groups suitable for subsequent bioorthogonal reactions and thus label RNA with reporter moieties such as affinity or fluorescent tags. In this review article, we will cover chemo‐enzymatic approaches (a) for in vitro labeling of RNA for application in cells, (b) for treatment of total RNA, and (c) for metabolic labeling of RNA. This article is categorized under: RNA Processing < RNA Editing and Modification RNA Methods < RNA Analyses in vitro and In Silico RNA Methods < RNA Analyses in Cells Understanding how RNA molecules orchestrate gene expression pathways and malfunction in disease requires investigations of localization, interactions, and dynamics of RNA in living cells. In this review article, we highlight chemo‐enzymatic methods developed to make RNA accessible to analyses. In particular,chemo‐enzymatic strategies for RNA labeling, enrichment and detection of intrinsic modifications will be presented.