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Gene introduction into mouse blastocysts via "pricking"
1Laboratory for Pathobiology, Pharma Research Laboratories, Hoechst Japan Ltd., Saitama.
Molecular Reproduction and Development
|April 1, 1993
Summary
Mouse blastocysts can be efficiently transfected with foreign DNA using a micromanipulation pricking technique. This method allows for gene expression in the inner cell mass (ICM), offering new avenues for genetic research in early development.
Area of Science:
- Developmental Biology
- Molecular Biology
- Genetics
Background:
- Mammalian cell transformation via pricking in the presence of foreign DNA is established.
- The potential for DNA uptake and gene expression in early mouse embryos remains an area for investigation.
Purpose of the Study:
- To investigate the efficiency of exogenous DNA uptake in mouse blastocysts using micromanipulation.
- To determine if introduced genes can be expressed within the blastocyst inner cell mass (ICM).
Main Methods:
- Mouse blastocysts were subjected to micromanipulation 'pricking' in the presence of linearized plasmid DNA.
- Polymerase chain reaction (PCR)-Southern analysis was used to detect foreign DNA in resulting fetuses.
- Beta-galactosidase (beta-gal) gene expression was assessed histochemically in cultured blastocysts.
Main Results:
- Fifty percent of blastocysts transferred to pseudopregnant females resulted in developing fetuses.
- Seventy-three percent of these fetuses carried foreign DNA, detected via PCR-Southern analysis.
- At least 65% of cultured blastocysts showed beta-gal activity, indicating successful gene expression in the ICM.
Conclusions:
- Micromanipulation pricking is an efficient method for transfecting mouse blastocysts with exogenous DNA.
- Introduced genes are expressed in the blastocyst ICM, validating the technique for genetic studies.
- This method can aid in mapping regulatory elements and studying ICM cell fate in vivo.