Use of Target Protector Morpholinos to Analyze the Physiological Roles of Specific MiRNA-mRNA Pairs in Vivo

Overview

Journal Nat Protoc

Specialties Biology
Pathology
Science

Date 2011 Dec 3

PMID 22134127

Citations 55

Authors

Alison A Staton

Antonio J Giraldez

Affiliations

Soon will be listed here.

Abstract

MicroRNAs (miRNAs) regulate gene expression by pairing with complementary sequences in the 3' 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 3' 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.

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