Cell type–specific manipulation with GFP-dependent Cre recombinase

• Published in: Nature neuroscience Vol. 18; no. 9; pp. 1334 - 1341
• Main Authors: Tang, Jonathan C Y, Rudolph, Stephanie, Dhande, Onkar S, Abraira, Victoria E, Choi, Seungwon, Lapan, Sylvain W, Drew, Iain R, Drokhlyansky, Eugene, Huberman, Andrew D, Regehr, Wade G, Cepko, Constance L
• Format: Journal Article
• Language: English
• Published: New York Nature Publishing Group US 01.09.2015; Nature Publishing Group
• Subjects: 631/1647/1511; 631/1647/2253; 631/378/3920; 64; 64/110; 82; Adeno-associated virus; Animal Genetics and Genomics; Animals; Behavioral Sciences; Biological Techniques; Biomedicine; Female; Gene expression; Genes; Genetic aspects; Genetic engineering; Green fluorescent protein; Green Fluorescent Proteins - analysis; HEK293 Cells; Humans; Integrases - analysis; Integrases - biosynthesis; Male; Medical schools; Mice; Mice, Inbred C57BL; Mice, Transgenic; Nervous system; Neurobiology; Neurons; Neurons - chemistry; Neurons - metabolism; Neurosciences; Ophthalmology; Optogenetics - methods; Organ Culture Techniques; Physiological aspects; Pregnancy; Proteins; Recombinases; Retina - chemistry; Retina - cytology; Retina - metabolism; technical-report; Transcription factors; United States; United States > US; Massachusetts; California
• ISSN: 1097-6256; 1546-1726
• DOI: 10.1038/nn.4081

GFP reporter lines are useful for labeling specific cell types. Here, the authors developed a method to convert GFP expression directly into Cre recombinase activity. GFP-dependent Cre was delivered via electroporation or AAV to neural tissues in the mouse, and could be used for optogenetic control of specific cell types. There are many transgenic GFP reporter lines that allow the visualization of specific populations of cells. Using such lines for functional studies requires a method that transforms GFP into a molecule that enables genetic manipulation. We developed a method that exploits GFP for gene manipulation, Cre recombinase dependent on GFP (CRE-DOG), a split component system that uses GFP and its derivatives to directly induce Cre/ loxP recombination. Using plasmid electroporation and AAV viral vectors, we delivered CRE-DOG to multiple GFP mouse lines, which led to effective recombination selectively in GFP-labeled cells. Furthermore, CRE-DOG enabled optogenetic control of these neurons. Beyond providing a new set of tools for manipulation of gene expression selectively in GFP + cells, we found that GFP can be used to reconstitute the activity of a protein not known to have a modular structure, suggesting that this strategy might be applicable to a wide range of proteins.