Format

Send to

Choose Destination
Nat Neurosci. 2015 Sep;18(9):1334-41. doi: 10.1038/nn.4081. Epub 2015 Aug 10.

Cell type-specific manipulation with GFP-dependent Cre recombinase.

Author information

1
Howard Hughes Medical Institute, Harvard Medical School, Boston, Massachusetts, USA.
2
Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.
3
Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts, USA.
4
Department of Neurobiology, Harvard Medical School, Boston, Massachusetts, USA.
5
Department of Neurosciences, University of California, San Diego, California, USA.
6
Neurobiology Section in the Division of Biological Sciences, University of California, San Diego, California, USA.
7
Department of Ophthalmology, University of California, San Diego, California, USA.

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.

PMID:
26258682
PMCID:
PMC4839275
DOI:
10.1038/nn.4081
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Nature Publishing Group Icon for PubMed Central
Loading ...
Support Center