Protocol for the neonatal intracerebroventricular delivery of adeno-associated viral vectors for brain restoration of MECP2 for Rett syndrome

Kan Yang; Tianshu Li; Yixiao Geng; Ru Zhang; Zhankui Xu; Jun Wu; Yiting Yuan; Yuefang Zhang; Zilong Qiu; Fei Li

doi:10.1016/j.xpro.2024.103344

Protocol for the neonatal intracerebroventricular delivery of adeno-associated viral vectors for brain restoration of MECP2 for Rett syndrome

STAR Protoc. 2024 Dec 20;5(4):103344. doi: 10.1016/j.xpro.2024.103344. Epub 2024 Sep 25.

Authors

Kan Yang¹, Tianshu Li², Yixiao Geng², Ru Zhang³, Zhankui Xu³, Jun Wu², Yiting Yuan², Yuefang Zhang², Zilong Qiu⁴, Fei Li⁵

Affiliations

¹ Department of Developmental and Behavioural Pediatric & Child Primary Care, Brain and Behavioural Research Unit of Shanghai Institute for Pediatric Research and MOE-Shanghai Key Laboratory for Children's Environmental Health of Xinhua Hospital & Songjiang Research Institute, Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China; College of Materials and Chemical Engineering, Hunan Institute of Engineering, Xiangtan, Hunan 411104 , China. Electronic address: kanyang@shsmu.edu.cn.
² Department of Developmental and Behavioural Pediatric & Child Primary Care, Brain and Behavioural Research Unit of Shanghai Institute for Pediatric Research and MOE-Shanghai Key Laboratory for Children's Environmental Health of Xinhua Hospital & Songjiang Research Institute, Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China.
³ College of Materials and Chemical Engineering, Hunan Institute of Engineering, Xiangtan, Hunan 411104 , China.
⁴ Department of Developmental and Behavioural Pediatric & Child Primary Care, Brain and Behavioural Research Unit of Shanghai Institute for Pediatric Research and MOE-Shanghai Key Laboratory for Children's Environmental Health of Xinhua Hospital & Songjiang Research Institute, Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China. Electronic address: qiuzilong@shsmu.edu.cn.
⁵ Department of Developmental and Behavioural Pediatric & Child Primary Care, Brain and Behavioural Research Unit of Shanghai Institute for Pediatric Research and MOE-Shanghai Key Laboratory for Children's Environmental Health of Xinhua Hospital & Songjiang Research Institute, Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China. Electronic address: feili@shsmu.edu.cn.

Abstract

Here, we present a protocol for neonatal intracerebroventricular (ICV) delivery of adeno-associated viral vectors (AAVs), achieving gene therapy for a Rett syndrome mouse model. We describe steps for preparing mouse lines, replacing foster mothers, sex typing, and genotyping. We then detail procedures for ICV delivery and validation through immunofluorescent and immunoblot techniques. This protocol is also applicable to preclinical gene therapy research that targets the neonatal mouse brain for other neurodevelopmental disorders. For complete details on the use and execution of this protocol, please refer to Yang et al.¹.

Keywords: clinical protocol; model organisms; neuroscience.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Animals, Newborn*
Brain* / metabolism
Dependovirus* / genetics
Disease Models, Animal*
Female
Gene Transfer Techniques
Genetic Therapy* / methods
Genetic Vectors* / administration & dosage
Genetic Vectors* / genetics
Injections, Intraventricular
Male
Methyl-CpG-Binding Protein 2* / genetics
Methyl-CpG-Binding Protein 2* / metabolism
Mice
Rett Syndrome* / genetics
Rett Syndrome* / therapy

Substances

Methyl-CpG-Binding Protein 2
Mecp2 protein, mouse