Anmelden

University of South China

3 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.