Shanghai Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine
2 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 Luo 4, Wen Xue 1,2, Xuelian Liu 1,2, Jiwu Wang 1,2,3, Ping Wei 4
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.
Biochemistry
CRISPR-based Shuttle Cloning: A High-throughput Cloning Method
Xiaoxue Li *1,2, Yutian Peng *2,3, Wen Xue 2, Xuelian Liu 2, Si Luo 4, Ping Wei 4, Jiwu Wang 2
1Institute of Biochemistry and Molecular Biology, Hengyang Medical School, University of South China, 2Clinical Research Institute, 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 describe a protocol for a high-throughput cloning method, CRISPR-based shuttle cloning (CRISPRshuttle cloning), which allows the transfer of DNA fragments of interest between vectors without the need for PCR amplification of the DNA fragments.