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Microinjection of dsRNA into mouse one-cell embryos
1Department of Biology, University of Pennsylvania, Philadelphia, PA 19104, USA. steinpau@sas.upenn.edu
Cold Spring Harbor Protocols
|February 12, 2010
Summary
This study details a method for gene silencing in mouse embryos using RNA interference (RNAi) via double-stranded RNA (dsRNA) microinjection. Researchers can assess gene knockdown efficiency using quantitative reverse transcriptase (RT)-PCR (qPCR).
Area of Science:
- Developmental Biology
- Molecular Biology
- Genetics
Background:
- RNA interference (RNAi) enables sequence-specific gene silencing.
- Effective delivery methods are crucial for applying RNAi in early embryonic development.
Purpose of the Study:
- To establish a protocol for delivering double-stranded RNA (dsRNA) into mouse one-cell embryos.
- To enable gene knockdown studies during early mouse development.
Main Methods:
- Microinjection of dsRNA into mouse one-cell embryos.
- Culture of microinjected embryos to the blastocyst stage (4 days).
- Assay of gene knockdown efficiency using quantitative reverse transcriptase (RT)-PCR (qPCR).
Main Results:
- Successful delivery of dsRNA into mouse embryos via microinjection.
- Embryos can be cultured to the blastocyst stage post-microinjection.
- Quantitative RT-PCR (qPCR) allows for efficient assessment of RNAi efficacy.
Conclusions:
- Microinjection of dsRNA is a viable method for RNA interference in mouse embryos.
- This protocol supports the study of gene function during early embryonic development.
- The method allows for efficient gene knockdown analysis using qPCR.
