Production and purification of endogenously modified tRNA-derived small RNAs

Abstract

2020): Production and purification of endogenously modified tRNA-derived small RNAs, RNA Biology, ABSTRACT During particular stress conditions, transfer RNAs (tRNAs) become substrates of stress-induced endonucleases, resulting in the production of distinct tRNA-derived small RNAs (tsRNAs). These small RNAs have been implicated in a wide range of biological processes, but how isoacceptor and even isodecoder-specific tsRNAs act at the molecular level is still poorly understood. Importantly, stress-induced tRNA cleavage affects only a few tRNAs of a given isoacceptor or isodecoder, raising the question as to how such limited molecule numbers could exert measurable biological impact. While the molecular function of individual tsRNAs is likely mediated through association with other molecules, addressing the interactome of specific tsRNAs has only been attempted by using synthetic RNA sequences. Since tRNAs carry post-transcriptional modifications, tsRNAs are likely modified but the extent of their modifications remains largely unknown. Here, we developed a biochemical framework for the production and purification of specific tsRNAs using human cells. Preparative scale purification of tsRNAs from biological sources should facilitate experimentally addressing as to how exactly these small RNAs mediate the multitude of reported molecular functions. ARTICLE HISTORY

Transfer RNA-Derived Small RNAs: Another Layer of Gene Regulation and Novel Targets for Disease Therapeutics

“…50,51,91,92,103 Currently, the purification methods of endogenous tsRNAs based on their modification state have just started to be developed. 124,125 More-advanced, modification-based tsRNA purification technology will be important for understanding the role of modification on tsRNA and developing tsRNAs as therapeutic drugs. Nevertheless, we have just started to scratch the surface in our understanding of the biological role of these non-coding RNAs.…”

Bacterial tRNA 2′-O-methylation is dynamically regulated under stress conditions and modulates innate immune response

“…The principle of individual tRNA isolation is outlined in the Supplementary Figure S1 . Pure tRNA isoacceptors were isolated by affinity column chromatography on covalently immobilized fully complementary synthetic DNA oligonucleotide ( 54 ). Further details are given in Supplementary Table S1 and in the Supplementary Data.…”

tsRNAs: Novel small molecules from cell function and regulatory mechanism to therapeutic targets

“…76 57 Theoretically, once the mode and location of the modification are determined, the expert laboratory can synthesize the modified nucleotides introduced at the specific location. 112 However, the current commercial use still needs further research and design.…”