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Generation of Adenovirus for In Vitro and In Vivo Studies of Hepatocytes.

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单位： [1]Cancer Center, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China. [2]Division of Endocrinology, Boston Children's Hospital, Harvard Medical School, Boston, MA, USA. [3]Division of Endocrinology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Branch of the National Clinical Research Center for Metabolic Disease, Wuhan, China. [4]Key Laboratory of Molecular Target & Clinical Pharmacology and the State Key Laboratory of Respiratory Disease, School of Pharmaceutical Sciences & the Fifth Affiliated Hospital, Guangzhou Medical University, Guangzhou, China. [5]Sapthagiri Institute of Medical Sciences and Research Centre, Bangalore, India. [6]Division of Endocrinology, Boston Children's Hospital, Harvard Medical School, Boston, MA, USA

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Although non-alcoholic steatohepatitis (NASH) can progress to liver cancer and liver failure, no FDA-approved drugs exist to treat NASH. Deciphering the molecular mechanisms underlying the pathogenesis of NASH will facilitate the development of effective treatments for NASH, and requires loss- or gain-of-function experimental approaches. While genetically modified animals provide important information about the function of a gene, adenovirus is a fast, effective, and versatile tool that allows transient knockdown, knockout, or overexpression of one or more genes of interest (GOIs) in primary hepatocytes in vitro and in mouse liver in vivo. In addition, adenovirus is a promising treatment method in preclinical animal models, including rodents and non-human primates, and is used in many clinical trials. Here, we describe a step-by-step protocol to generate adenovirus for basic medical research. We discuss critical steps during virus propagation and purification and provide notes about how to avoid common pitfalls.© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

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第一作者单位： [1]Cancer Center, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China. [2]Division of Endocrinology, Boston Children's Hospital, Harvard Medical School, Boston, MA, USA.

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