Format

Send to

Choose Destination
See comment in PubMed Commons below
PLoS One. 2010 Dec 16;5(12):e14375. doi: 10.1371/journal.pone.0014375.

A transient transgenic RNAi strategy for rapid characterization of gene function during embryonic development.

Author information

1
Genetics Division, Brigham & Women's Hospital, Harvard Medical School, Boston, Massachusetts, United States of America.

Abstract

RNA interference (RNAi) is a powerful strategy for studying the phenotypic consequences of reduced gene expression levels in model systems. To develop a method for the rapid characterization of the developmental consequences of gene dysregulation, we tested the use of RNAi for "transient transgenic" knockdown of mRNA in mouse embryos. These methods included lentiviral infection as well as transposition using the Sleeping Beauty (SB) and PiggyBac (PB) transposable element systems. This approach can be useful for phenotypic validation of putative mutant loci, as we demonstrate by confirming that knockdown of Prdm16 phenocopies the ENU-induced cleft palate (CP) mutant, csp1. This strategy is attractive as an alternative to gene targeting in embryonic stem cells, as it is simple and yields phenotypic information in a matter of weeks. Of the three methodologies tested, the PB transposon system produced high numbers of transgenic embryos with the expected phenotype, demonstrating its utility as a screening method.

PMID:
21179568
PMCID:
PMC3002952
DOI:
10.1371/journal.pone.0014375
[Indexed for MEDLINE]
Free PMC Article
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for Public Library of Science Icon for PubMed Central
    Loading ...
    Support Center