Evaluating the susceptibility of AGO2-loaded microRNAs to degradation by nucleases in vitro

Reyad Elbarbary, Lynne E. Maquat

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

MicroRNAs (miRNAs) comprise a class of small non-coding RNAs that regulate the stability and/or translatability of most protein-coding transcripts. Steady-state levels of mature miRNAs can be controlled through mechanisms that influence their biogenesis and/or decay rates. Pathways that mediate mature miRNA decay are less well understood than those that mediate miRNA biogenesis. We recently described Tudor-staphylococcal/micrococcal-like nuclease (TSN)-mediated miRNA decay (TumiD) as a cellular pathway that promotes the sequence-specific endonucleolytic decay of miRNAs that harbor a CA and/or UA dinucleotide. Here, we describe an in vitro assay for evaluating the susceptibility of AGO2-loaded miRNAs to degradation by different classes of nucleases. This in vitro approach can be used to complement in vivo studies that aim to identify novel miRNA decay factors.

Original languageEnglish (US)
Pages (from-to)18-22
Number of pages5
JournalMethods
Volume152
DOIs
StatePublished - Jan 1 2019

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MicroRNAs
Degradation
Micrococcal Nuclease
Small Untranslated RNA
In Vitro Techniques
RNA Stability
Ports and harbors
Assays
Proteins

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

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abstract = "MicroRNAs (miRNAs) comprise a class of small non-coding RNAs that regulate the stability and/or translatability of most protein-coding transcripts. Steady-state levels of mature miRNAs can be controlled through mechanisms that influence their biogenesis and/or decay rates. Pathways that mediate mature miRNA decay are less well understood than those that mediate miRNA biogenesis. We recently described Tudor-staphylococcal/micrococcal-like nuclease (TSN)-mediated miRNA decay (TumiD) as a cellular pathway that promotes the sequence-specific endonucleolytic decay of miRNAs that harbor a CA and/or UA dinucleotide. Here, we describe an in vitro assay for evaluating the susceptibility of AGO2-loaded miRNAs to degradation by different classes of nucleases. This in vitro approach can be used to complement in vivo studies that aim to identify novel miRNA decay factors.",
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Evaluating the susceptibility of AGO2-loaded microRNAs to degradation by nucleases in vitro. / Elbarbary, Reyad; Maquat, Lynne E.

In: Methods, Vol. 152, 01.01.2019, p. 18-22.

Research output: Contribution to journalArticle

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