Rapid Production and Dissemination of Mouse Intersectional Genetic Alleles for Neural Circuit Mapping
The goal of this project is to build and distribute a community library of more than 100 mouse intersectional genetic alleles to facilitate the developmental, anatomical, molecular, and functional characterization of neural circuit organization in behavior and physiology
Intersectional Genetic Allele Generation Pipeline
Flow chart for proposed intersectional genetic mouse pipeline. Production of mouse alleles will be guided by public input. All reagents generated including plasmids, ES cells, and mouse lines will be deposited in accessible public respositories for unrestricted access.
Intersectional Genetics
An example of intersectional genetics used to identify two subpopulations of pyramidal neurons in the cortex. A) The intersectional allele consists of four key elements: 1) a broadly active CAGpromoter; 2) an FRT flanked stop cassette; 3) a LoxPflanked mCherry and stop cassette and 4) an intersectional eGFP reporter. B) Delineation of cortical pyramidal neurons expressing Gene A::FLP and Gene B::Cre transgenes. The subset of neurons expressing Gene A will be marked with mCherry upon FLP recombination. The subset of Gene A neurons that also express Gene B will be marked with eGFP expression (intersectional) upon CRE mediated LoxP recombination, and leaving the remaining FLP (mCherry population) as the subtractive population. Importantly, those pyramidal neurons that express Gene B outside the context of Gene A will remain unmarked, since the first stop cassette remains in place.
Funding
This work is funded by generous support from the Office Of The Director, National Institutes Of Health (1R21OD025327)
