Rapid generation of gene-targeted EPS-derived mouse models through tetraploid complementation.

Area of Science:

• Stem cell biology
• Genetics
• Developmental biology

Background:

• Gene targeting in mouse embryonic stem (ES) cells is crucial for creating genetically modified mouse models.
• Long-term culture of ES cells leads to genetic and epigenetic instability, reducing germline transmission efficiency.
• Extended pluripotent stem (EPS) cells represent a novel cell type with enhanced developmental potency and germline competence.

Purpose of the Study:

• To evaluate the stability and developmental potency of mouse EPS cells after long-term culture.
• To establish a rapid method for generating gene-targeted mouse models using EPS cells and tetraploid complementation.
• To demonstrate the feasibility of creating specific gene-knocked mouse models using this novel approach.

Main Methods:

• Culturing and characterization of mouse extended pluripotent stem (EPS) cells.
• Gene targeting within EPS cells.
• Tetraploid complementation for rapid generation of gene-targeted mouse models.
• Generation of mouse models with human IL3 or IL6 gene knock-in.

Main Results:

• Mouse EPS cells maintained developmental potency and genetic stability during long-term passage.
• A rapid method for generating gene-targeted mouse models was established, taking approximately 2 months.
• Successfully generated mouse models with human IL3 or IL6 genes integrated into the mouse genome.

Conclusions:

• Mouse EPS cells are a stable and competent alternative to ES cells for generating genetically modified mouse models.
• Gene targeting in EPS cells combined with tetraploid complementation offers a significantly faster and more robust approach to mouse model generation.
• This method holds great potential for accelerating biomedical research by enabling rapid creation of custom mouse models.