PUBLICATION
Use of target protector morpholinos to analyze the physiological roles of specific miRNA-mRNA pairs in vivo
- Authors
- Staton, A.A., and Giraldez, A.J.
- ID
- ZDB-PUB-111205-14
- Date
- 2011
- Source
- Nature Protocols 6(12): 2035-2049 (Journal)
- Registered Authors
- Giraldez, Antonio, Staton, Alison
- Keywords
- genetic analysis, nucleic acid based molecular biology
- MeSH Terms
-
- 3' Untranslated Regions
- Animals
- Binding, Competitive
- Embryo, Nonmammalian
- Gene Expression Regulation
- Genetic Techniques
- Luciferases/analysis
- MicroRNAs/chemistry
- MicroRNAs/physiology*
- Morpholinos/chemistry*
- RNA, Messenger/chemistry
- RNA, Messenger/physiology*
- Zebrafish/embryology
- Zebrafish/genetics*
- PubMed
- 22134127 Full text @ Nat. Protoc.
Citation
Staton, A.A., and Giraldez, A.J. (2011) Use of target protector morpholinos to analyze the physiological roles of specific miRNA-mRNA pairs in vivo. Nature Protocols. 6(12):2035-2049.
Abstract
MicroRNAs (miRNAs) regulate gene expression by pairing with complementary sequences in the 32 untranslated regions (UTRs) of transcripts. Although the molecular mechanism underlying miRNA biogenesis and activity is becoming better understood, determining the physiological role of the interaction of an miRNA with its target remains a challenge. A number of methods have been developed to inhibit individual miRNAs, but it can be difficult to determine which specific targets are responsible for any observed phenotypes. To address this problem, we use target protector (TP) morpholinos that interfere with a single miRNA-mRNA pair by binding specifically to the miRNA target sequence in the 32 UTR. In this protocol, we detail the steps for identifying miRNA targets, validating their regulation and using TPs to interrogate their function in zebrafish. Depending on the biological context, this set of experiments can be completed in 6–8 weeks
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping