University of South China

4 ARTICLES PUBLISHED IN JoVE

Genetics

CRISPR-Based Modular Assembly for High-Throughput Construction of a UAS-cDNA/ORF Plasmid Library

Si Xu^*1,2, Mengyu Chen^*1,2, Guang Chen^1,3, Si Luo⁴, Wen Xue^1,2, Xuelian Liu^1,2, Jiwu Wang^1,2,3, Ping Wei⁴

¹Clinical Research Institute, The Affiliated Nanhua Hospital, Hengyang Medical School, University of South China, ²Department of Neurology, The Affiliated Nanhua Hospital, Hengyang Medical School, University of South China, ³Department of Clinical Laboratory, The Affiliated Nanhua Hospital, Hengyang Medical School, University of South China, ⁴Shanghai Diabetes Institute, Shanghai Key Laboratory of Diabetes Mellitus, Shanghai Clinical Center for Diabetes, Shanghai Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine

We present a protocol for CRISPR-based modular assembly (CRISPRmass), a method for high-throughput construction of UAS-cDNA/ORF plasmid library in Drosophila using publicly available cDNA/ORF resources. CRISPRmass can be applied to editing various plasmid libraries.

Genetics

Embryo Microinjection for Transgenesis in Drosophila

Guang Chen^*1,2, Xinyi Zhang^*1,3, Si Xu^1,4, Xiaotao Zhou¹, Wen Xue¹, Xuelian Liu¹, Jialong Yan¹, Nana Zhang¹, Jiwu Wang^1,2,4

¹Clinical Research Institute, The Affiliated Nanhua Hospital, Hengyang Medical School, University of South China, ²Department of Clinical Laboratory, The Affiliated Nanhua Hospital, Hengyang Medical School, University of South China, ³Institute of Biochemistry and Molecular Biology, Hengyang Medical School, University of South China, ⁴Department of Neurology, The Affiliated Nanhua Hospital, Hengyang Medical School, University of South China

This article takes the phiC31 integrase-mediated transgenesis in Drosophila as an example and presents an optimized protocol for embryo microinjection, a crucial step for creating transgenic flies.

Cancer Research

Proximity Ligation Assay to Study Oncogene-Derived Transcription-Replication Conflicts

Linling Ke¹, Qian Xie¹, Xiaoman Wang¹, Yuanhang Gong¹, Min Li^1,2

¹Institute of Biochemistry and Molecular Biology, Hengyang Medical School, University of South China, ²National Health Commission Key Laboratory of Birth Defect Research and Preventio, Hunan Provincial Maternal and Child Health Care Hospital

Here, we present a sensitive and specific method to detect oncogene-induced transcription-replication conflicts (TRCs) using the proximity ligation assay (PLA). This approach leverages specific antibodies targeting PCNA and phospho-CTD RNAPII, enabling the assessment of TRC prevalence between RNA polymerase II transcription and DNA replication machinery.

Immunology and Infection

Development and Assessment of Intracellular Infection Models for Staphylococcus aureus

Chengwen Zhang^1,2, Chuanfei Xu², Fumei Luo^2,3, Yu Zuo², Ping Luo², Ping Cheng², Weijun Zhang², Si Sun², Hao Zeng², Quanming Zou²

¹Medical Research Institute, Southwest University, ²National Engineering Research Center of Immunological Products, Third Military Medical University, ³School of Pharmacy, University of South China

Staphylococcus aureus (S. aureus) has the capability to disseminate throughout the body, causing persistent and recurrent infections. To better understand these processes, this study establishes an intracellular infection model for S. aureus. This model will provide a crucial foundation for investigating the mechanisms behind intracellular infections.