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

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. 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. 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 ( ). 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.