A CRISPR toolbox for generating intersectional genetic mouse models for functional, molecular, and anatomical circuit...

Area of Science:

• Genetics
• Molecular Biology
• Animal Models

Background:

• Understanding cellular mechanisms in health and disease requires studying discrete cell subtypes.
• Recombinase-driven expression is a powerful method for delineating cell populations.
• Current methods for creating intersectional mouse lines are expensive and technically challenging.

Purpose of the Study:

• To present a simplified CRISPR toolkit for rapid, inexpensive, and facile intersectional allele production.
• To overcome limitations of existing methods for creating customized animal models.

Main Methods:

• Developed a CRISPR toolkit for producing intersectional alleles.
• Generated 7 intersectional mouse lines using a dual recombinase system.
• Created 1 single recombinase system mouse line and 3 embryonic stem (ES) cell lines.

Main Results:

• Successfully produced diverse, modifiable animal models using the genetic toolkit.
• Applied new mouse lines for anatomical mapping and functional investigation of neuronal populations controlling respiratory control.
• Utilized a single recombinase-responsive allele to study hormone effects on calcium release in ovarian follicles.

Conclusions:

• The toolkit facilitates the easy creation of single or dual recombinase effector lines for any cell population of interest.
• Openly available tools and cell lines support non-commercial research.
• This approach enables the development of diverse, modifiable animal models for various research applications.