ESC-Based Gene Editing

The Embryonic Stem Cell Targeting Process in Mice

Embryonic stem cell or ESC-based gene targeting uses a stem cell’s plasticity to engineer gene targeted mice. Mouse stem cells are first electroporated with a targeting vector. Following successful homologous recombination between the targeting vector and ESC cell DNA, targeted embryonic stem cells are then injected into a normal mouse blastocyst. When injected, the stem cells colonize the blastocyst, resulting in a chimera with cells of distinct genotypes. Blastocysts are then implanted into a host mouse that will carry and deliver it. Finally, chimeric (founder) mice are bred with strains of mice to produce animals that have germline transmission of the targeted gene.

Advantages of Biocytogen’s ESC-based gene editing

Although traditional way to edit genomes but ESC/HR based gene editing offers following advantages:

Service Workflow

Knockout Mouse Models

In ESC-based gene knockout targeting, we use a positive selection marker to replace an exon of a targeted gene. The targeted gene is deleted globally, whereas in conditional knockout animals the gene is deleted only in selected cells, as determined by a tissue specific promoter. Conditional knockout mouse models are usually generated using Cre-LoxP or Flp-Frt recombination systems.

Knockin Mouse Models

To generate a knockin mouse, a DNA fragment of interest is inserted at the desired location in the genome. This modality also allows for a variety of models to be created including point mutant, reporter (GFP), tagged (FLAG), Cre, and humanized mice.

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