Cite as: Cold Spring Harb. Protoc.; 2009; doi:10.1101/pdb.prot5135

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Microinjection of Plasmids into Meiotically Incompetent 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 mammalian double-stranded RNA (dsRNA) expression vector into meiotically incompetent mouse oocytes by microinjection. During mouse oocyte growth, there is a progressive decline in transcriptional activity, such that the fully-grown oocyte is transcriptionally quiescent. For this reason, the microinjection of plasmids should be performed in growing oocytes, obtained from 12- to 13-d-old females. Microinjected oocytes can be cultured for up to 2 d. The efficiency of knockdown by RNAi can be assayed by quantitative reverse transcriptase (RT)-PCR (qPCR).


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RNAi Experiments in Mouse Oocytes and Early Embryos
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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
Cloning a Transgene for Transgenic RNAi in Mouse Oocytes
CSH Protocols, January 1, 2009; 2009(1): pdb.prot5134 - pdb.prot5134.
[Abstract] [Full Text]