FLP-mediated site-specific recombination in microinjected murine zygotes

Area of Science:

• Molecular Biology
• Genetics
• Biotechnology

Background:

• The FLP recombinase enzyme from yeast facilitates site-specific DNA recombination.
• This recombination occurs between specific DNA sequences known as FLP recombinase target (FRT) sites.
• FLP recombinase has been shown to function in various organisms and cell types.

Purpose of the Study:

• To investigate the efficacy of FLP recombinase in manipulating the genome of early-stage mouse embryos.
• To determine if FLP recombinase can activate reporter genes and modify chromosomal DNA in microinjected mouse eggs.

Main Methods:

• Transient expression of FLP recombinase was achieved by coinjecting it into fertilized one-cell mouse eggs.
• A FLP-recombinase expression vector was introduced into transgenic one-cell fertilized mouse eggs.
• Reporter gene activation and chromosomal recombination events were analyzed.

Main Results:

• Transient FLP recombinase expression successfully recombined and activated an extrachromosomal reporter gene in mouse eggs.
• Introduction of a FLP-recombinase vector induced recombination at a chromosomal FRT target locus in the one-cell stage.
• This chromosomal event resulted in the deletion of a tandem array containing an FRT-lacZ cassette.

Conclusions:

• The FLP recombinase is a viable tool for genome manipulation in transgenic animals.
• FLP recombinase-mediated targeting of plasmid DNA to chromosomal loci is feasible in microinjected mouse eggs.
• This technology holds potential for genetic engineering in early embryonic development.