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Development of a Corneal Bioluminescence Mouse for Real-Time In Vivo Evaluation of Gene Therapies.
Fu, Dun Jack; Allen, Edwin H A; Hickerson, Robyn P; Leslie Pedrioli, Deena M; McLean, W H Irwin.
Afiliación
  • Fu DJ; Centre for Dermatology and Genetic Medicine, Division of Biological Chemistry and Drug Discovery, School of Life Sciences, University of Dundee, Dundee, Scotland, UK.
  • Allen EHA; Centre for Dermatology and Genetic Medicine, Division of Biological Chemistry and Drug Discovery, School of Life Sciences, University of Dundee, Dundee, Scotland, UK.
  • Hickerson RP; School of Biomedical Sciences, University of Ulster, Coleraine, Northern Ireland, UK.
  • Leslie Pedrioli DM; Centre for Dermatology and Genetic Medicine, Division of Biological Chemistry and Drug Discovery, School of Life Sciences, University of Dundee, Dundee, Scotland, UK.
  • McLean WHI; Centre for Dermatology and Genetic Medicine, Division of Biological Chemistry and Drug Discovery, School of Life Sciences, University of Dundee, Dundee, Scotland, UK.
Transl Vis Sci Technol ; 9(13): 44, 2020 12.
Article en En | MEDLINE | ID: mdl-33442498
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 (luc2) was achieved via insertion into the Krt12 locus, a type I intermediate filament keratin that is exclusively expressed in the cornea, to generate the Krt12luc2 mouse. The transgene includes a multiple target cassette with human pathogenic mutations in K3 and K12. Results: The Krt12luc2 mouse exclusively expresses luc2 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 luc2 transgene (Krt12+/luc2). Conclusions: This novel Krt12luc2 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 Luc2 transgene will greatly reduce mouse numbers required for in vivo therapy evaluation.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Epitelio Corneal / Enfermedades de la Córnea Límite: Animals Idioma: En Revista: Transl Vis Sci Technol Año: 2020 Tipo del documento: Article Pais de publicación: Estados Unidos
Texto completo
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Imprimir
XML
PubMed Links
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Epitelio Corneal / Enfermedades de la Córnea Límite: Animals Idioma: En Revista: Transl Vis Sci Technol Año: 2020 Tipo del documento: Article Pais de publicación: Estados Unidos