Xiao Dan, Zhang Weifeng, Wang Qing, Li Xing, Zhang Yuan, Rasouli Javad, Casella Giacomo, Ciric Bogoljub, Curtis Mark, Rostami Abdolmohamad, Zhang Guang-Xian
Department of Neurology, Thomas Jefferson University, Philadelphia, PA 19107, USA.
College of Life Sciences, Shaanxi Normal University, Xi'an, Shaanxi 710062, China.
Mol Ther Methods Clin Dev. 2021 Feb 18;20:755-764. doi: 10.1016/j.omtm.2021.02.012. eCollection 2021 Mar 12.
Inducible conditional knockout mice are important tools for studying gene function and disease therapy, but their generation is costly and time-consuming. We introduced clustered regularly interspaced short palindromic repeats (CRISPR) and into an LSL-Cas9 transgene-carrying mouse line by using adeno-associated virus (AAV)-PHP.eB to rapidly knockout gene(s) specifically in central nervous system (CNS) cells of adult mice. in neurons and in astrocytes were knocked out 2 weeks after an intravenous injection of vector, with an efficiency comparable to that of inducible Cre- conditional knockout. For functional testing, we generated astrocyte-specific knockout mice, which exhibited a phenotype similar to mice with Cre--mediated knockout, in an animal model of multiple sclerosis (MS), an autoimmune disorder of the CNS. With this novel technique, neural cell-specific knockout can be induced rapidly (few weeks) and cost-effectively. Our study provides a new approach to building inducible conditional knockout mice, which would greatly facilitate research on CNS biology and disease.
诱导性条件性基因敲除小鼠是研究基因功能和疾病治疗的重要工具，但其构建成本高且耗时。我们通过使用腺相关病毒（AAV）-PHP.eB将成簇规律间隔短回文重复序列（CRISPR）引入携带LSL-Cas9转基因的小鼠品系，以快速在成年小鼠的中枢神经系统（CNS）细胞中特异性敲除基因。静脉注射载体2周后，神经元中的[具体基因1]和星形胶质细胞中的[具体基因2]被敲除，其效率与诱导性Cre条件性基因敲除相当。为了进行功能测试，我们构建了星形胶质细胞特异性[具体基因3]敲除小鼠，在中枢神经系统自身免疫性疾病多发性硬化症（MS）的动物模型中，其表现出与Cre介导的[具体基因3]敲除小鼠相似的表型。通过这种新技术，可以快速（几周内）且经济高效地诱导神经细胞特异性基因敲除。我们的研究为构建诱导性条件性基因敲除小鼠提供了一种新方法，这将极大地促进中枢神经系统生物学和疾病的研究。
