PUBLICATION
Performing the Labeled microRNA Pull-Down (LAMP) Assay System: An Experimental Approach for High-Throughput Identification of microRNA-Target mRNAs
- Authors
- Hsu, R.J., and Tsai, H.J.
- ID
- ZDB-PUB-110713-68
- 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.
Citation
Hsu, R.J., and Tsai, H.J. (2011) Performing the Labeled microRNA Pull-Down (LAMP) Assay System: An Experimental Approach for High-Throughput Identification of microRNA-Target mRNAs. Methods in molecular biology (Clifton, N.J.). 764:241-247.
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping