A Publicly Guided Effort to Rapidly Produce and Disseminate 100 Intersectional Genetic Alleles for the Study of Nervous System Circuit Development and Function in the Mouse

Abstract

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 based on public input. Even within narrowly defined cell types, significant diversity is found at multiple levels including genetic and molecular signatures, activity patterns, and synaptic connectivity. Current challenges now center on tools to identify, access, and study cell populations with increasing precision. Intersectional genetics offers exceptionally high resolution to consistently delineate distinct cell types in the embryo and adult mouse for functional, molecular and anatomical studies. Intersectional genetics utilizes a ubiquitously expressed conditional allele that is activated by Cre and Flp site specific recombinases. Upon activation, the intersectional genetic alleles may express any number of Genetically Encodable Effector Molecules (GEEMS), such as channelrhodopsins for neural activity perturbations or an L10‐GFP ribotrap fusion to affinity purify translating mRNAs. The intersectional allele is activated by overlapping expression of both Flp and Cre recombinases in the same cell. Traditionally, these recombinases have been deployed as gene knock‐in or transgenic alleles that are designed to express in a cell or gene specific fashion. The use of two selectors (Cre and Flp) to define a cellular population enables high specificity and modularity in combining any set of Cre, Flp and intersectional alleles to fit an experiment. The number of Cre and Flp recombinase mouse lines are constantly growing, giving greater access to increasing numbers of cell types. Additionally, these recombinases are the focus of multiple efforts to deploy them in ways that select cells based on other unique properties such as neuronal activity and synaptic connectivity. To make this powerful genetic approach more accessible to as many research laboratories as possible, we are building a production pipeline to create a suite of resources for anyone to easily make their own intersectional mouse line, to produce over 100 targeted ES cell lines that can be developed into mouse lines, as well as 10–15 high demand mouse lines all for public distribution. Priority for the GEEMS to be intersectionally deployed will be based on a public input collected at mouseintersectionalgenetics.org. All plasmids, cell lines and mouse lines developed under this effort will be distributed without restriction to non‐profit research organizations.Support or Funding InformationMcNair Medical InstituteR01HL130249R01HL130249‐02S1This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.