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Updated: Jun 28, 2025
Limbal Approach-Subretinal Injection of Viral Vectors for Gene Therapy in Mice Retinal Pigment Epithelium
Published on: August 7, 2015
Intraocular mRNA delivery with endogenous MmPEG10-based virus-like particles
Mengke Li1, Zhong Liu2, Dongliang Wang2
1State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangdong Provincial Key Laboratory of Ophthalmology and Visual Science, Guangzhou, 510060, China; Research Unit of Ocular Development and Regeneration, Chinese Academy of Medical Sciences, Beijing, 100085 China.
New virus-like particles (VLPs) effectively deliver mRNA to eye cells, including retinal pigment epithelial and corneal cells. These VLPs show promise for ocular gene therapy without causing significant immune responses.
Area of Science:
- Ocular gene therapy
- Molecular and cell biology
- Biotechnology
Background:
- Virus-like particles (VLPs) are potent intracellular gene delivery tools.
- Their application in ocular gene therapy is largely unexplored.
- Addressing this gap is crucial for advancing eye disease treatments.
Purpose of the Study:
- To generate and evaluate Vesicular Stomatitis Virus Glycoprotein (VSVG)-pseudotyped mouse PEG10 (MmPEG10)-VLP for ocular mRNA delivery.
- To assess the efficacy of MmPEG10-VLP in transducing retinal pigment epithelial (RPE) and corneal cells.
- To investigate the potential of MmPEG10-VLP for therapeutic gene transfer, such as SMAD7 delivery.
Main Methods:
- Generation of VSVG-pseudotyped MmPEG10-VLP.
- In vitro delivery of GFP mRNA to ARPE-19 cells.
- In vivo subretinal and intravitreal injections in adult mice.
- Intracameral delivery to assess corneal transduction.
- Evaluation of immune response and therapeutic gene transfer (SMAD7).
Main Results:
- PEG10-VLP efficiently delivered GFP mRNA to ARPE-19 cells, resulting in transient expression.
- MmPEG10-VLP successfully transferred SMAD7, inhibiting epithelial-mesenchymal transition in RPE cells.
- Subretinal delivery in mice led to efficient RPE transduction and GFP expression without significant immune response.
- Intracameral delivery showed transient GFP expression in corneal endothelial cells.
- Intravitreal injection was inefficient for ocular expression.
Conclusions:
- VSVG-pseudotyped MmPEG10-based VLPs are effective for transducing mitotically inactive RPE cells and corneal endothelial cells in vivo.
- These VLPs demonstrate potential for ocular gene therapy applications, including gene transfer to RPE and corneal cells.
- The lack of significant inflammatory response highlights the safety profile of these VLPs for ocular delivery.
