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Background  Genetic targeting methods have greatly advanced our understanding of many of the 20 Retinal Ganglion Cell (RGC) types conveying visual information from the eyes to the brain. However, the complexity and partial overlap of gene expression patterns in RGCs call for genetic intersectional or sparse labeling strategies. Loci carrying the Cre recombinase in conjunction with conditional knock-out, reporter or other genetic tools can be used for targeted cell type ablation and functional manipulation of specific cell populations. The three members of the Pou4f family of transcription factors, Brn3a, Brn3b and Brn3c, expressed early during RGC development and in combinatorial pattern amongst RGC types are excellent candidates for such gene manipulations.    Methods and Findings  We generated conditional Cre knock-in alleles at the Brn3a and Brn3b loci,  Brn3a CKOCre   and  Brn3b CKOCre  . When crossed to mice expressing the Dre recombinase, the endogenous Brn3 gene expressed by  Brn3a CKOCre   or  Brn3b CKOCre   is removed and replaced with a Cre recombinase, generating  Brn3a Cre   and  Brn3b Cre   knock-in alleles. Surprisingly both  Brn3a Cre   and  Brn3b Cre   knock-in alleles induce early ubiquitous recombination, consistent with germline expression. However in later stages of development, their expression is limited to the expected endogenous pattern of the  Brn3a  and  Brn3b  genes. We use the  Brn3a Cre   and  Brn3b Cre   alleles to target a Cre dependent Adeno Associated Virus (AAV) reporter to RGCs and demonstrate its use in morphological characterization, early postnatal gene delivery and tracing the expression of Brn3 genes in RGCs.    Conclusions  Dre recombinase effectively recombines the  Brn3a CKOCre   and  Brn3b CKOCre   alleles containing its roxP target sites. Sequential Dre to Cre recombination reveals Brn3a and Brn3b expression in early mouse development. The generated  Brn3a Cre   and  Brn3b Cre   alleles are useful tools that can target exogenously delivered Cre dependent reagents to RGCs in early postnatal development, opening up a large range of potential applications.

Dre - Cre Sequential Recombination Provides New Tools for Retinal Ganglion Cell Labeling and Manipulation in Mice