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Tudor C Badea, Nadia Parmhans, Eileen Nguyen, Katherine Chuang; Combining Dre and Cre recombination to identify and study mouse Retinal Ganglion Cell Types. Invest. Ophthalmol. Vis. Sci. 2017;58(8):1749.
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© ARVO (1962-2015); The Authors (2016-present)
We are designing mouse genetic strategies for the labeling and manipulation of specific Retinal Ganglion Cell Types. Previously we have reported the use of sequential Dre and Cre recombination employing the genetic loci of the Pou4f transcription factors to isolate individual cell populations. To improve the temporal specificity and tighten the Dre-dependency of Cre expression, we identified novel heterotopic Dre sites, and designed inversion-excision strategies targeting the Cre recombinase at the Pou4f3 locus.
Targeting constructs, ES cell targeting by homologous recombination and blastocyst injections were performed by established methodologies. The generated mouse lines were tested by crossing into available Dre and Cre and reporter lines.
We report successful Dre to Cre to Cre reporter recombination resulting in RGC specific expression of the reporter. We provide neuroanatomic and molecular characterization of the lines, and apply them to RGC type characterization.
Gene expression profiling of neuronal cell types provides ample evidence of the broad overlap of gene expression between neuronal cell types. Therefore, genetic intersection strategies are crucial for the specific isolation of individual cell types. Strategies employing more than one recombinase can be successfully applied for specific cell type manipulation in live, otherwise intact animals.
This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.
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