3 articles published in JoVE
CRISPR/Cas9-mediated Targeted Integration In Vivo Using a Homology-mediated End Joining-based Strategy Xuan Yao1,2, Xing Wang1,2, Junlai Liu2,3,4, Linyu Shi1, Pengyu Huang3, Hui Yang1 1Institute of Neuroscience, State Key Laboratory of Neuroscience, Key Laboratory of Primate Neurobiology, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 2College of Life Sciences, University of Chinese Academy of Sciences, 3School of Life Science and Technology, Shanghai Tech University, 4Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences The clustered regularly interspaced short palindromic repeats/CRISPR associated protein 9 (CRISPR/Cas9) system provides a promising tool for genetic engineering, and opens up the possibility of targeted integration of transgenes. We describe a homology-mediated end joining (HMEJ)-based strategy for efficient DNA targeted integration in vivo and targeted gene therapies using CRISPR/Cas9.
Whole-cell Patch-clamp Recordings of Isolated Primary Epithelial Cells from the Epididymis Bao Li Zhang1,2,3, Da Yuan Gao1,2,3, Xiao Xu Zhang1,3, Shuo Shi4, Winnie Shum1 1School of Life Science and Technology, ShanghaiTech University, 2Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 3University of Chinese Academy of Sciences, 4Shanghai Institute for Advanced Immunochemical Studies, ShanghaiTech University We present a protocol that combines cell isolation and whole-cell patch-clamp recording to measure the electrical properties of the primary dissociated epithelial cells from the rat cauda epididymides. This protocol allows for investigation of the functional properties of primary epididymal epithelial cells to further elucidate the physiological role of the epididymis.
Production of Chemicals by Klebsiella pneumoniae Using Bamboo Hydrolysate as Feedstock Dong Wei1,2, Jinjie Gu1, Zhongxi Zhang1, Chenhong Wang1, Dexin Wang1, Chul Ho Kim3, Biao Jiang1, Jiping Shi1,4, Jian Hao1 1Lab of Biorefinery, Shanghai Advanced Research Institute, Chinese Academy of Sciences, 2University of Chinese Academy of Sciences, 3Biorefinery Research Center, Jeonbuk Branch Institute, Korea Research Institute of Bioscience & Biotechnology (KRIBB), 4School of Life Science and Technology, ShanghaiTech University Bamboo powder was pretreated with NaOH and enzymatically hydrolyzed. The hydrolysate of bamboo was used as the feedstock for 2,3-butanediol, R-acetoin, 2-ketogluconic acid, and xylonic acid production by Klebsiella pneumoniae.