alexa Evaluation of AAV-Mediated Gene Therapy with Reduced Vector Volume in Cngb3 Knockout Mice, a Model of Achromatopsia
ISSN: 2161-1041

Hereditary Genetics: Current Research
Open Access

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Research Article

Evaluation of AAV-Mediated Gene Therapy with Reduced Vector Volume in Cngb3 Knockout Mice, a Model of Achromatopsia

Xuan Liu1, Yuxin Zhang1,2, Wei Du1, Wei Shi1, Ye Tao1, Wen-Tao Deng1, Jie Li1, Chen Zhao2 and Ji-jing Pang1,2,3*

1Department of Ophthalmology, College of Medicine, University of Florida, Gainesville, Florida

2Department of Ophthalmology, First Affiliated Hospital, Nanjing Medical University, Nanjing, Jiangsu, China

3Eye Hospital, School of Ophthalmology and Optometry, Wenzhou Medical University, Wenzhou, Zhejiang, China

*Corresponding Author:
Ji-jing Pang
Department of Ophthalmology
College of Medicine
University of Florida
Gainesville, Florida, USA
Tel: 352-273-9341
E-mail: [email protected]

Received date: November 24, 2015; Accepted date: February 23, 2016; Published date: February 26, 2016

Citation: Liu X, Zhang Y, Du W, Shi W, Tao Y, et al. (2016) Evaluation of AAVMediated Gene Therapy with Reduced Vector Volume in Cngb3 Knockout Mice, a Model of Achromatopsia. Hereditary Genet 5:163. doi:10.4172/2161-1041.1000163

Copyright: © 2016 Liu X, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

 

Abstract

Purpose: This study was designed to investigate whether the volume of vector used for subretinal injection can be reduced to transfect C57bl/6J mouse whole retina and whether it can restore cone function in a Cngb3 knockout (KO) mouse model.
Methods: C57bl/6J mice and Cngb3 KO mice received a subretinal injection of 0.5 μL or 1 μL of AAV5-smCBAmCherry vector and AAV5-IRBP/GNAT2-hCngb3 vector, respectively. Retinal whole mounts and frozen sections were prepared from the wild-type mouse eyes to evaluate the transfected area. Dark and light-adapted electroretinograms (ERGs) were recorded two months after vector injection in the eyes of Cngb3 KO mice.
Result: In the retina of AAV5-smCBA-mCherry injected wild-type mice, no difference was observed between the injection volumes. mCherry positive retinal pigment epithelial (RPE) and photoreceptor cells were observed throughout the entire retina. In AAV5-IRBP/GNAT2-hCngb3-injected Cngb3 KO mice, 1-μL-injected mice showed a higher average of photopic ERG restoration than 0.5-μL-injected mice. However, the scotopic ERGs were lower in 1-μL-injected mice, indicating that higher injection volumes resulted in more damages.
Conclusion: Reduced volume (0.5 μL) of vector induced fewer damages. However, higher doses of vector (1 μL) restore higher ERG function in Cngb3 KO mouse.

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