基于伪狂犬病病毒巴塔株的Cre依赖型逆行跨多突触示踪剂的研发。
Development of Cre-dependent retrograde trans-multisynaptic tracer based on pseudorabies virus bartha strain.
作者信息
You Hu, Qinghan Wang, Kangyixin Sun, Jia Yang, Fuqiang Xu, Fan Jia
机构信息
Guangdong Provincial Key Laboratory of Brain Connectome and Behavior, CAS Key Laboratory of Brain Connectome and Manipulation, the Brain Cognition and Brain Disease Institute, Shenzhen Institutes of Advance Technology, Translational Research Center for the Nervous System, Chinese Academy of Sciences, Shenzhen-Hong Kong Institute of Brain Science-Shenzhen Fundamental Research Institutions, Shenzhen, 518055, China.
NMPA Key Laboratory for Research and Evaluation of Viral Vector Technology in Cell and Gene Therapy Medicinal Products, Key Laboratory of Quality Control Technology for Virus-Based Therapeutics, Shenzhen Key Laboratory of Viral Vectors for Biomedicine, Shenzhen Institutes of Advance Technology, Guangdong Provincial Medical Products Administration, Chinese Academy of Sciences, Shenzhen, 518055, China.
出版信息
Mol Brain. 2025 Apr 14;18(1):33. doi: 10.1186/s13041-025-01204-y.
Mapping the neural circuit of a specific neuronal subclass is central to understanding the working mechanism of the brain. Currently, numerous types of transgenic mice expressing Cre recombinase have been engineered and widely used in neuroscience. To map the multilevel inputs into the neural circuit of a specific neuronal subpopulation, a Cre-dependent retrograde trans-multisynaptic tracer must be developed. The vaccine strain of Pseudorabies virus (PRV, Bartha strain) can infect neurons and spread in a retrograde manner in the neural circuit. In this study, we engineered the genome of PRV Bartha strain to prepare two new tracers, PRV676 and PRV829, by replacing the TK gene of PRV with the Cre-dependent expression cassette of the fluorescent protein gene and the TK gene. These two tracers can separately and Cre-dependently express EGFP and mRuby3 and produce progeny viruses in vitro and in vivo, which can help to map the multilevel inputs of a specific neuronal subpopulation expressing Cre. Collectively, our work provides two new tools for neuroscience research.
绘制特定神经元亚类的神经回路对于理解大脑的工作机制至关重要。目前，已经构建了多种表达Cre重组酶的转基因小鼠，并在神经科学中广泛应用。为了绘制特定神经元亚群神经回路的多级输入，必须开发一种Cre依赖性逆行跨多突触示踪剂。伪狂犬病病毒（PRV，Bartha株）疫苗株可感染神经元并在神经回路中逆行传播。在本研究中，我们通过用荧光蛋白基因的Cre依赖性表达盒和TK基因替换PRV的TK基因，对PRV Bartha株的基因组进行改造，制备了两种新的示踪剂PRV676和PRV829。这两种示踪剂可以分别以Cre依赖性方式表达EGFP和mRuby3，并在体外和体内产生子代病毒，这有助于绘制表达Cre的特定神经元亚群的多级输入。总的来说，我们的工作为神经科学研究提供了两种新工具。
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