Authors:
Jonathan C Y Tang, Stephanie Rudolph, Onkar S Dhande, Victoria E Abraira, Seungwon Choi, Sylvain W Lapan, Iain R Drew, Eugene Drokhlyansky, Andrew D Huberman, Wade G Regehr, & Constance L Cepko
Summary:
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.
Source:
Nature Neuroscience; (08/10/15)