ZFIN ID: ZDB-PUB-110713-68
Performing the Labeled microRNA Pull-Down (LAMP) Assay System: An Experimental Approach for High-Throughput Identification of microRNA-Target mRNAs
Hsu, R.J., and Tsai, H.J.
Date: 2011
Source: Methods in molecular biology (Clifton, N.J.)   764: 241-247 (Chapter)
Registered Authors: Tsai, Huai-Jen
Keywords: none
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*
PubMed: 21748645 Full text @ Meth. Mol. Biol.
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.