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RESEARCH PRODUCT
Absolute quantification of noncoding RNA by microscale thermophoresis
Raffael SchaffrathYuri MotorinKathrin ThüringMark HelmMichael StockRoland KlassenAkif CiftciVirginie MarchandSebastian A. LeidelJean-yves RoignantAurellia GalliotAdeline GalvaninKarin ScharmannDominik Jacobsubject
tRNA stabilityRNA UntranslatedAbsolute quantificationRNA Quantification | Hot PaperComputational biology010402 general chemistry01 natural sciencesCatalysis[SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry Molecular Biology/Genomics [q-bio.GN]RNA modification540 ChemistryhybridizationComputingMilieux_MISCELLANEOUS010405 organic chemistryChemistryMicroscale thermophoresisCommunicationRNA[SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry Molecular Biology/Molecular biologyGeneral ChemistryRibosomal RNANon-coding RNAmicroscale thermophoresisCommunications0104 chemical sciencesTissue DifferentiationTransfer RNA570 Life sciences; biologyfluorescenceRNA quantificationdescription
Abstract Accurate quantification of the copy numbers of noncoding RNA has recently emerged as an urgent problem, with impact on fields such as RNA modification research, tissue differentiation, and others. Herein, we present a hybridization‐based approach that uses microscale thermophoresis (MST) as a very fast and highly precise readout to quantify, for example, single tRNA species with a turnaround time of about one hour. We developed MST to quantify the effect of tRNA toxins and of heat stress and RNA modification on single tRNA species. A comparative analysis also revealed significant differences to RNA‐Seq‐based quantification approaches, strongly suggesting a bias due to tRNA modifications in the latter. Further applications include the quantification of rRNA as well as of polyA levels in cellular RNA.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 2019-03-20 |