Proteomics

Dataset Information

0

A universal method for the rapid isolation of all known classes of functional small RNAs

ABSTRACT: Diverse classes of regulatory small (s)RNAs operate via ARGONAUTE-family proteins within RNA-induced-silencing-complexes (RISCs). Based the conserved biochemical properties intrinsic to all ARGONAUTEs, we have developed a universal, 15-min benchtop extraction procedure allowing simultaneous purification of all classes of RISC-associated sRNAs, without prior knowledge of the sample’s -intrinsic ARGONAUTE repertoires. Optimized as a user-friendly kit, the method –coined “TraPR” for Trans-kingdom, rapid, affordable Purification of RISCs– operates irrespectively of the organism, tissue, cell type or bio-fluid of interest, and scales to minute amounts of input material. The method is highly suited for direct sRNA deep-sequencing, with TraPR-generated libraries being qualitatively at least on-par with those obtained via gold-standard procedures that require immunoprecipitations and/or lengthy polyacrylamide gel excisions. TraPR considerably improves the quality and consistency of sRNA sample preparation including from notoriously difficult-to-handle tissues/bio-fluids such as starchy storage roots and mammalian plasma, and regardless of RNA contaminants or samples’ RNA-degradation status.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Caenorhabditis Elegans

SUBMITTER: Julia Svozil  

LAB HEAD: Olivier Voinnet

PROVIDER: PXD015395 | Pride | 2021-09-08

REPOSITORIES: Pride

Json Xml
altmetric image

Publications

A universal method for the rapid isolation of all known classes of functional silencing small RNAs.

Grentzinger Thomas T   Oberlin Stefan S   Schott Gregory G   Handler Dominik D   Svozil Julia J   Barragan-Borrero Veronica V   Humbert Adeline A   Duharcourt Sandra S   Brennecke Julius J   Voinnet Olivier O  

Nucleic acids research 20200801 14

Diverse classes of silencing small (s)RNAs operate via ARGONAUTE-family proteins within RNA-induced-silencing-complexes (RISCs). Here, we have streamlined various embodiments of a Q-sepharose-based RISC-purification method that relies on conserved biochemical properties of all ARGONAUTEs. We show, in multiple benchmarking assays, that the resulting 15-min benchtop extraction procedure allows simultaneous purification of all known classes of RISC-associated sRNAs without prior knowledge of the sa  ...[more]

Similar Datasets

Proteomics GFPtrap enrichment of CYP83A1 and CYP83B1
2017-11-30 | PXD005585 | Pride
Proteomics Protein abundance changes and ubiquitylation targets identified upon inhibition of the proteasome with Syringolin A
2014-04-15 | PXD000521 | Pride
Proteomics Protein abundance changes and ubiquitylation targets identified upon inhibition of the proteasome with Syringolin A
2014-04-15 | PXD000566 | Pride
Proteomics Differences in proteasome targeting in Arabidopsis leaf mesophyll, epidermal and vascular cells
2015-05-14 | PXD000941 | Pride
Proteomics Protein abundance changes and ubiquitylation targets identified upon inhibition of the proteasome with Syringolin A
2014-04-15 | PXD000567 | Pride
Proteomics Protein abundance changes and ubiquitylation targets identified upon inhibition of the proteasome with Syringolin A
2014-05-19 | PXD000568 | Pride
Proteomics Protein abundance changes and ubiquitylation targets identified upon inhibition of the proteasome with Syringolin A
2014-04-15 | PXD000565 | Pride
Proteomics Differences in proteasome targeting in Arabidopsis leaf mesophyll, epidermal and vascular cells
2015-05-13 | PXD000942 | Pride
Transcriptomics Coupling between alternative polyadenylation and alternative splicing is limited to terminal introns
2016-03-14 | E-GEOD-79157 | biostudies-arrayexpress
Unknown A universal, benchtop, gel-free method for the rapid and simultaneous isolation of all known classes of functional small RNAs
2020-07-01 | GSE134516 | GEO