Cite as: Cold Spring Harb. Protoc.; 2009; doi:10.1101/pdb.prot5132

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Microinjection of dsRNA into Fully-Grown Mouse Oocytes

Paula Stein

Department of Biology, University of Pennsylvania, Philadelphia, PA 19104, USA

Corresponding author (steinpau{at}sas.upenn.edu)


INTRODUCTION

RNA interference (RNAi) is a suitable method for sequence-specific post-transcriptional gene silencing in a number of model systems. The following protocol describes delivery of a double-stranded RNA (dsRNA) of choice into fully-grown, germinal vesicle-intact (GV) mouse oocytes by microinjection. Microinjected oocytes can be cultured for up to 2 d or they can be matured to metaphase II. The metaphase II eggs can be fertilized in vitro and cultured up to the blastocyst stage. The efficiency of knockdown by RNAi can be assayed by quantitative reverse transcriptase (RT)-PCR (qPCR).


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Related Article

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This article has been cited by other articles:


Home page
 CSH ProtocolsHome page
P. Svoboda and P. Stein
RNAi Experiments in Mouse Oocytes and Early Embryos
CSH Protocols, January 1, 2009; 2009(1): pdb.top56 - pdb.top56.
[Abstract] [Full Text]

Home page
 CSH ProtocolsHome page
P. Svoboda
Preparation of dsRNA for Microinjection Experiments in Mouse
CSH Protocols, January 1, 2009; 2009(1): pdb.prot5131 - pdb.prot5131.
[Abstract] [Full Text]