Knock-in human rhodopsin-GFP fusions as mouse models for human disease and targets for gene therapy.

Abstract

The human rhodopsin gene is the locus for numerous alleles linked to the neurodegenerative disease retinitis pigmentosa. To facilitate the study of retinal degeneration and to test reagents designed to alter the structure and function of this gene, we have developed strains of mice whose native rhodopsin gene has been replaced with the corresponding human DNA modified to encode an enhanced GFP fusion at the C terminus of rhodopsin. The human rhodopsin-GFP fusion faithfully mimics the expression and distribution of wild-type rhodopsin in heterozygotes and serves as a sensitive reporter of rod-cell structure and integrity. In homozygotes, however, the gene induces progressive retinal degeneration bearing many of the hallmarks of recessive retinitis pigmentosa. When the gene is flanked by recognition sites for Cre recombinase, protein expression is reduced approximately 5-fold despite undiminished mRNA levels, suggesting translation inhibition. GFP-tagged human rhodopsin provides a sensitive method to monitor the development of normal and diseased retinas in dissected samples, and it offers a noninvasive means to observe the progress of retinal degeneration and the efficacy of gene-based therapies in whole animals.

• Authors: Fung Chan, Allan Bradley, Theodore G Wensel, John H Wilson
• Journal: Proceedings of the National Academy of Sciences of the United States of America (Proc Natl Acad Sci U S A) Vol. 101 Issue 24 Pg. 9109-14 (Jun 15 2004) ISSN: 0027-8424 [Print] United States
• PMID: 15184660 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Luminescent Proteins
• Recombinant Fusion Proteins
• Green Fluorescent Proteins
• Rhodopsin

Topics

• Animals
• Animals, Genetically Modified
• Blastocyst (metabolism)
• Cell Nucleus (metabolism)
• Cell Survival
• Disease Models, Animal
• Gene Expression
• Genetic Therapy (methods)
• Green Fluorescent Proteins
• Humans
• Luminescent Proteins (genetics, metabolism)
• Male
• Mice
• Recombinant Fusion Proteins (genetics, metabolism)
• Retina (cytology, metabolism, pathology, ultrastructure)
• Retinal Degeneration (genetics, metabolism)
• Rhodopsin (genetics, metabolism)
• Stem Cells (metabolism)