Practical considerations for analyzing antigene RNAs (agRNAs): RNA immunoprecipitation of argonaute protein

Jacob C. Schwartz, David R. Corey

Research output: Chapter in Book/Report/Conference proceedingChapter

3 Scopus citations

Abstract

Target validation for small RNAs in cells can be a confusing task wrought with pitfalls and false leads. One technique for validating in vivo targets of small RNAs is immunoprecipitation of target RNAs using antibodies again the RNAi machinery. Antigene RNAs (agRNAs) regulate transcription in human cells using machinery from the RNAi regulatory pathway-namely argonaute proteins. Here we describe a technique for validating targets of agRNAs using RNA immunoprecipitation with antibodies against human argonaute proteins. This technique can be used to detect interactions of argonaute proteins in the cell nucleus with their targets, lowly expressed noncoding RNA transcripts.

Original languageEnglish (US)
Title of host publicationTherapeutic Oligonucleotides
Subtitle of host publicationMethods and Protocols
EditorsJohn Goodchild
Pages301-315
Number of pages15
DOIs
StatePublished - 2011

Publication series

NameMethods in Molecular Biology
Volume764
ISSN (Print)1064-3745

Keywords

  • Noncoding RNA
  • RNA immunoprecipitation
  • antigene RNAs
  • argonaute
  • transcriptional gene silencing

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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