Development of a Corneal Bioluminescence Mouse for Real-Time In Vivo Evaluation of Gene Therapies.

Purpose: The purpose of this study was to develop and characterize a novel bioluminescence transgenic mouse model that facilitates rapid evaluation of genetic medicine delivery methods for inherited and acquired corneal diseases.

Methods: Corneal expression of the firefly luciferase transgene () was achieved via insertion into the locus, a type I intermediate filament keratin that is exclusively expressed in the cornea, to generate the mouse. The transgene includes a multiple target cassette with human pathogenic mutations in K3 and K12.

Results: The mouse exclusively expresses in the corneal epithelium under control of the keratin K12 promoter. The luc2 protein is enzymatically active, can be readily visualized, and exhibits a symmetrically consistent readout. Moreover, structural integrity of the corneal epithelium is preserved in mice that are heterozygous for the transgene ().

Conclusions: This novel mouse model represents a potentially ideal in vivo system for evaluating the efficacies of cornea-targeting gene therapies and for establishing and/or validating new delivery modalities. Importantly, the multiple targeting cassette that is included in the transgene will greatly reduce mouse numbers required for in vivo therapy evaluation.