Work

Cre-assisted fine-mapping of neural circuits using orthogonal split inteins

Year: 2020

Type: article

Abstract: Existing genetic methods of neuronal targeting do not routinely achieve the resolution required for mapping brain circuits. New approaches are thus necessary. Here, we introduce a method for refined neuronal targeting that can be applied iteratively. Restriction achieved at the first step can be further refined in a second step, if necessary. The method relies on first isolating neurons within a targeted group (i.e. Gal4 pattern) according to their developmental lineages, and then intersectionally limiting the number of lineages by selecting only those in which two distinct neuroblast enhancers are active. The neuroblast enhancers drive expression of split Cre recombinase fragments. These are fused to non-interacting pairs of split inteins, which ensure reconstitution of active Cre when all fragments are expressed in the same neuroblast. Active Cre renders all neuroblast-derived cells in a lineage permissive for Gal4 activity. We demonstrate how this system can facilitate neural circuit-mapping in Drosophila.

Source: eLife

DOI: 10.7554/elife.53041

Authors: Haojiang Luan, Alexander Kuzin, Ward F. Odenwald, Benjamin H. White

Institutions: National Institutes of Health, National Institute of Mental Health, National Institute of Neurological Disorders and Stroke

Language: English

Topic: Neurobiology and Insect Physiology Research

Subfield: Cellular and Molecular Neuroscience

Field: Neuroscience

Domain: Life Sciences

Open Access status: gold

Estimated APC paid: $2,500

Funders: National Institutes of Health, National Institute of Mental Health, National Institute of Neurological Disorders and Stroke