Limbal Approach-Subretinal Injection of Viral Vectors for Gene Therapy in Mice Retinal Pigment Epithelium

The eye is a small and enclosed organ which makes it an ideal target for gene therapy. Recently various strategies have been applied to gene therapy in retinopathies using non-viral and viral gene delivery to the retina and retinal pigment epithelium (RPE). Subretinal injection is the best approach...

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Bibliographic Details
Published in:Journal of visualized experiments 2015-08 (102), p.e53030-e53030
Main Authors: Park, Sung Wook, Kim, Jin Hyoung, Park, Woo Jin, Kim, Jeong Hun
Format: Article
Language:English
Subjects:
Adenoviridae - genetics
Animals
Dependovirus - genetics
Genetic Therapy - methods
Genetic Vectors - administration & dosage
Green Fluorescent Proteins - administration & dosage
Green Fluorescent Proteins - biosynthesis
Green Fluorescent Proteins - genetics
Lentivirus - genetics
Limbus Corneae
Luminescent Proteins - administration & dosage
Luminescent Proteins - biosynthesis
Luminescent Proteins - genetics
Mice
Neuroscience
Red Fluorescent Protein
Retinal Diseases - genetics
Retinal Diseases - therapy
Retinal Pigment Epithelium
Transduction, Genetic - methods
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Summary:The eye is a small and enclosed organ which makes it an ideal target for gene therapy. Recently various strategies have been applied to gene therapy in retinopathies using non-viral and viral gene delivery to the retina and retinal pigment epithelium (RPE). Subretinal injection is the best approach to deliver viral vectors directly to RPE cells. Before the clinical trial of a gene therapy, it is inevitable to validate the efficacy of the therapy in animal models of various retinopathies. Thus, subretinal injection in mice becomes a fundamental technique for an ocular gene therapy. In this protocol, we provide the easy and replicable technique for subretinal injection of viral vectors to experimental mice. This technique is modified from the intravitreal injection, which is widely used technique in ophthalmology clinics. The representative results of RPE/choroid/scleral complex flat-mount will help to understand the efficacy of this technique and adjust the volume and titer of viral vectors for the extent of gene transduction.
ISSN:1940-087X
1940-087X
DOI:10.3791/53030