Zhengzhou University

6 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

A Simple and Efficient Approach to Construct Mutant Vaccinia Virus Vectors

Ming Yuan^*1, Pengju Wang^*2,3, Louisa S. Chard¹, Nicholas R. Lemoine^1,2,3, Yaohe Wang^1,2,3

¹Center for Molecular Oncology, Barts Cancer Institute, Queen Mary University of London, ²Sino-British Research Centre for Molecular Oncology, National Center for International Research in Cell and Gene Therapy, Zhengzhou University, ³School of Basic Medical Sciences, Academy of Medical Sciences, Zhengzhou University

Vaccinia virus (VV) has been widely used in biomedical research and the improvement of human health. This article describes a simple, highly efficient method to edit the VV genome using a CRISPR-Cas9 system.

Bioengineering

Production of Genetically Engineered Golden Syrian Hamsters by Pronuclear Injection of the CRISPR/Cas9 Complex

Rong Li^*1, Jinxin Miao^*1,2, Zhiqiang Fan¹, SeokHwan Song³, Il-keun Kong^3,4, Yaohe Wang^2,5, Zhongde Wang¹

¹Department of Animal, Dairy, and Veterinary Sciences, Utah State University, ²National Centre for International Research in Cell and Gene Therapy, Sino-British Research Centre, School of Basic Medical Sciences, Zhengzhou University, ³Department of Animal Science Division of Applied Life Science (BK21 Plus), Gyeongsang National University, ⁴Institute of Agriculture and Life Science, Gyeongsang National University, ⁵Centre for Molecular Oncology, Barts Cancer Institute, Queen Mary University of London

Pronuclear (PN) injection of the clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein-9 nuclease (CRISPR/Cas9) system is a highly efficient method for producing genetically engineered golden Syrian hamsters. Herein, we describe the detailed PN injection protocol for the production of gene knockout hamsters with the CRISPR/Cas9 system.

Bioengineering

Protocols of 3D Bioprinting of Gelatin Methacryloyl Hydrogel Based Bioinks

Mingjun Xie^1,2, Kang Yu^1,2, Yuan Sun^1,2, Lei Shao^1,2, Jing Nie^1,2, Qing Gao^1,2, Jingjiang Qiu³, Jianzhong Fu^1,2, Zichen Chen^1,2, Yong He^1,2

¹State Key Laboratory of Fluid Power and Mechatronic Systems, School of Mechanical Engineering, Zhejiang University, ²Key Laboratory of 3D Printing Process and Equipment of Zhejiang Province, School of Mechanical Engineering, Zhejiang University, ³School of Mechanics and Safety Engineering, Zhengzhou University

Presented here is a method for the 3D bioprinting of gelatin methacryloyl.

Immunology and Infection

In Vivo Augmentation of Gut-Homing Regulatory T Cell Induction

Hongzheng Bi^1,2, Samiksha Wasnik¹, David J. Baylink¹, Chenfan Liu^1,3,4, Xiaolei Tang^1,3

¹Division of Regenerative Medicine, Department of Medicine, Loma Linda University, ²Zhengzhou University, ³Department of Veterinary Biomedical Sciences, College of Veterinary Medicine, Long Island University, ⁴Jinan Infectious Disease Hospital, Shandong University

Here we present a protocol for in vivo augmentation of gut-homing regulatory T cell induction. In this protocol, dendritic cells are engineered to locally produce high concentrations of the active vitamin D (1,25-dihydroxyvitamin D or 1,25[OH]₂D) and the active vitamin A (retinoic acid or RA) de novo.

JoVE Core

Quantitative Polymerase Chain Reaction (qPCR)-Based Rapid Diagnosis of Helicobacter pylori Infection and Antibiotic Resistance

Liang Wang^*1,2,3, Jin-Xin Lai^*1, Yu-Ting Si^*1,4, Xu-Xia Cui^1,4, Zeeshan Umar^1,5, Xiao-Jun Ru¹, Xin-Yu Zhang¹, Zheng-Kang Li¹, Alfred Chin Yen Tay^5,6,7,8, Barry J. Marshall^5,6,7,8, Guang-Hua Li¹, Bing Gu¹

¹Laboratory Medicine, Guangdong Provincial People’s Hospital (Guangdong Academy of Medical Sciences), Southern Medical University, ²Division of Microbiology and Immunology, School of Biomedical Sciences, The University of Western Australia, ³Center for Precision Health, School of Medical and Health Sciences, Edith Cowan University, ⁴Medical Technology School of Xuzhou Medical University, ⁵Marshall Laboratory of Biomedical Engineering, School of Medicine, Shenzhen University, ⁶The Marshall Centre for Infectious Diseases Research and Training, The University of Western Australia, ⁷Marshall International Digestive Diseases Hospital, Zhengzhou University, ⁸Marshall Medical Research Center, Fifth Affiliated Hospital of Zhengzhou University

The protocol presents a noninvasive method for the rapid diagnosis of Helicobacter pylori stomach infections through the string test and determines its antibiotic resistance to clarithromycin and levofloxacin using quantitative polymerase chain reaction (qPCR).

Neuroscience

Measurement of Mitochondrial Membrane Potential In Vivo using a Genetically Encoded Voltage Indicator

Run-Zhou Yang¹, Dian-Dian Wang^2,3, Sen-Miao Li^2,3, Pei-Pei Liu¹, Jian-Sheng Kang^1,2,3

¹Clinical Systems Biology Laboratories, The First Affiliated Hospital of Zhengzhou University, ²Department of Neurology, The First Affiliated Hospital of Zhengzhou University, ³The Academy of Medical Sciences, Zhengzhou University

This protocol describes the application of mitochondria-targeted genetically encoded voltage indicators (GEVIs). These GEVIs offer a significant advantage over traditional mitochondrial membrane potential dyes by enabling specific, in vivo, and real-time monitoring of mitochondrial membrane potential.