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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.<\/jats:p>","DOI":"10.7554\/elife.53041","type":"journal-article","created":{"date-parts":[[2020,4,14]],"date-time":"2020-04-14T08:00:29Z","timestamp":1586851229000},"source":"Crossref","is-referenced-by-count":7,"title":["Cre-assisted fine-mapping of neural circuits using orthogonal split inteins"],"prefix":"10.7554","volume":"9","author":[{"given":"Haojiang","family":"Luan","sequence":"first","affiliation":[{"name":"Laboratory of Molecular Biology, National Institute of Mental Health, NIH, Bethesda, United States"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alexander","family":"Kuzin","sequence":"additional","affiliation":[{"name":"Neural Cell-Fate Determinants Section, National Institute of Neurological Disorders and Stroke, NIH, Bethesda, United States"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ward 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