Cell type–specific manipulation with GFP-dependent Cre recombinase

Abstract

Summary There are many transgenic GFP reporter lines that allow 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. Here we report the creation of 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, leading to effective recombination selectively in GFP-labeled cells. Further, 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 demonstrate 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.

Cell type- and stage-specific expression of Otx2 is coordinated by a cohort of transcription factors and multiplecis-regulatory modules in the retina

“…The Stagia3 vector was from Billings (Billings et al 2010) . The CAG-BFP plasmid was from Tang (Tang et al 2015) . All other plasmids were constructed by restrictive enzyme-based cloning or Gibson assembly methods (Gibson et al 2009).…”

Intrinsic functional and architectonic heterogeneity of tumor-targeted protein nanoparticles

“…Being functional but also biocompatible and biodegradable, protein materials show a still unexplored biomedical potential in both regenerative medicine and conventional or cell-targeted drug delivery [12,13]. The natural tendency of GFP to oligomerize [14] and the more recent manipulation of GFP assembling [15,16] have attracted interest as this beta-sheet rich protein represents a compact, structurally stable building block for the assay of controlled oligomer formation and material characterization.…”

Ipsilateral and Contralateral Retinal Ganglion Cells Express Distinct Genes during Decussation at the Optic Chiasm

“…The vertebrate retina, with its three cellular layers and six neuronal classes, has been a useful model for studying general principles of neurogenesis and axon guidance. Each class of retinal cells can be further divided into morphologically and functionally distinct subtypes, and recent efforts have identified the molecular programs that establish these differences within neuronal classes, such as amacrine, bipolar, and retinal ganglion cell (RGC) subtypes (Kim et al, 2008; Badea et al, 2009; Kay et al, 2011a, 2011b; Watson et al, 2012; Jiang et al, 2013; Sajgo et al, 2014; Macosko et al, 2015; Osterhout et al, 2015; Sanes and Masland, 2015; Tang et al, 2015; Jin et al, 2015; Rousso et al, 2016; Shekhar et al, 2016). RGCs, as the only projection neurons of the retina, can be additionally distinguished by the laterality of their axonal projection to targets in the thalamus and midbrain.…”