Cell type-specific manipulation with GFP-dependent Cre recombinase

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

Research output: Contribution to journalArticlepeer-review

35 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)1334-1341
Number of pages8
JournalNature Neuroscience
Volume18
Issue number9
DOIs
StatePublished - Aug 26 2015
Externally publishedYes

ASJC Scopus subject areas

  • Neuroscience(all)

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