Performing the Labeled microRNA pull-down (LAMP) assay system: an experimental approach for high-throughput identification of microRNA-target mRNAs

Methods Mol Biol. 2011;764:241-7. doi: 10.1007/978-1-61779-188-8_16.

Abstract

We developed a simple, direct, and cost-effective approach to search for the most likely target genes of a known miRNA in vitro. We term this method "Labeled microRNA (miRNA) pull-down assay system," or LAMP. Briefly, the pre-miRNA is labeled with digoxigenin (DIG), mixed with cell extracts, and immunoprecipitated by anti-DIG antiserum. We concluded that LAMP is an experimental approach for high-throughput identification of the target gene of known miRNAs from both Caenorhabditis elegans and zebrafish (Danio rerio), yielding fewer false-positive results than those produced by using only the bioinformatics approach.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Caenorhabditis elegans / genetics*
  • Digoxigenin / chemistry
  • Digoxigenin / immunology
  • High-Throughput Screening Assays / methods*
  • Immune Sera / chemistry
  • Immune Sera / immunology
  • Immunoprecipitation / methods*
  • MicroRNAs* / analysis
  • MicroRNAs* / genetics
  • MicroRNAs* / isolation & purification
  • RNA, Messenger / antagonists & inhibitors
  • RNA, Messenger / genetics
  • Zebrafish / genetics*

Substances

  • Immune Sera
  • MicroRNAs
  • RNA, Messenger
  • Digoxigenin