title: Cell type-specific manipulation with GFP-dependent Cre recombinase.
abstract: 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.
authors.affiliation: Howard Hughes Medical Institute, Harvard Medical School, Boston, Massachusetts, USA.|Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.|Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: Jonathan C Y
authors.initials: JC
authors.lastname: Tang
authors.affiliation: Department of Neurobiology, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Rudolph
authors.affiliation: Department of Neurosciences, University of California, San Diego, California, USA.|Neurobiology Section in the Division of Biological Sciences, University of California, San Diego, California, USA.|Department of Ophthalmology, University of California, San Diego, California, USA.
authors.forename: Onkar S
authors.initials: OS
authors.lastname: Dhande
authors.affiliation: Department of Neurobiology, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: Victoria E
authors.initials: VE
authors.lastname: Abraira
authors.affiliation: Department of Neurobiology, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: Seungwon
authors.initials: S
authors.lastname: Choi
authors.affiliation: Howard Hughes Medical Institute, Harvard Medical School, Boston, Massachusetts, USA.|Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.|Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: Sylvain W
authors.initials: SW
authors.lastname: Lapan
authors.affiliation: Department of Neurobiology, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: Iain R
authors.initials: IR
authors.lastname: Drew
authors.affiliation: Howard Hughes Medical Institute, Harvard Medical School, Boston, Massachusetts, USA.|Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.|Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: Eugene
authors.initials: E
authors.lastname: Drokhlyansky
authors.affiliation: Department of Neurosciences, University of California, San Diego, California, USA.|Neurobiology Section in the Division of Biological Sciences, University of California, San Diego, California, USA.|Department of Ophthalmology, University of California, San Diego, California, USA.
authors.forename: Andrew D
authors.initials: AD
authors.lastname: Huberman
authors.affiliation: Department of Neurobiology, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: Wade G
authors.initials: WG
authors.lastname: Regehr
authors.affiliation: Howard Hughes Medical Institute, Harvard Medical School, Boston, Massachusetts, USA.|Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.|Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: Constance L
authors.initials: CL
authors.lastname: Cepko
mesh_terms: D000818:Animals; D005260:Female; D049452:Green Fluorescent Proteins / Q000032:analysis*; D057809:HEK293 Cells; D006801:Humans; D019426:Integrases / Q000032:analysis* / Q000096:biosynthesis; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009474:Neurons / Q000737:chemistry* / Q000378:metabolism; D062308:Optogenetics / Q000379:methods*; D009924:Organ Culture Techniques; D011247:Pregnancy; D012160:Retina / Q000737:chemistry* / Q000166:cytology* / Q000378:metabolism
chemical_list: D049452:Green Fluorescent Proteins; C045073:Cre recombinase; D019426:Integrases