Quantification of microRNAs, splicing isoforms, and homologous mRNAs with the invader assay

Peggy S. Eis, Mariano A. Garcia-Blanco

Research output: Chapter in Book/Report/Conference proceedingChapter

3 Scopus citations


The understanding of physiology and pathology requires accurate quantification of intracellular concentrations of important molecules such as unique RNA species. Accurate quantification of highly homologous messenger RNAs (mRNAs) (1-3), alternatively spliced mRNAs (4), and the short microRNAs (miRNAs) (5,6) has been successfully achieved using the Invader assay. This method directly detects specific RNA molecules in preparations of pure total cellular RNA (1- 100 ng) or in crude cell lysate (103-104 cells) samples using an isothermal signal amplification process with a fluorescence resonance energy transfer (FRET)-based fluorescence readout. Features of the Invader assay include the ability to detect 1-10 RNA molecules per cell, to discriminate between RNAs that differ by a single base, and to precisely measure 1.2-fold changes in RNA expression. Further, an isothermal format and the ability to detect two different RNA molecules with a biplex format make the Invader assay suitable for high-throughput screening applications.

Original languageEnglish (US)
Title of host publicationRNA-Protein Interaction Protocols
PublisherHumana Press
Number of pages40
ISBN (Print)9781588294197
StatePublished - 2008
Externally publishedYes

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745


  • Alternative splicing
  • Cleavase enzyme
  • FRET
  • Gene expression
  • HTS
  • High-throughput screening
  • Invader assay
  • Invasive cleavage
  • RNA quantification/quantitation
  • Splice variant
  • mRNA
  • miRNA
  • microRNA

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics


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