Fujian Province University 2 articles published in JoVE Biology Generation of Centromere-Associated Protein-E CENP-E-/- Knockout Cell Lines using the CRISPR/Cas9 System Meng-Fei Xu1,2, Jie Chen1,2, Yue Xu1,2, Jing-Lian Zhang1,2, Yi Zhou1,2, Jie-Jie He1,2, Shan Wu1,2, Ya-Lan Wei3,4, Zhen-Yu She1,2 1Department of Cell Biology and Genetics, The School of Basic Medical Sciences, Fujian Medical University, 2Key Laboratory of Stem Cell Engineering and Regenerative Medicine, Fujian Province University, 3College of Clinical Medicine for Obstetrics & Gynecology and Pediatrics, Fujian Medical University, 4Medical Research Center, Fujian Maternity and Child Health Hospital This article reports the construction of centromere-associated protein-E (CENP-E) knockout cells using the CRISPR/Cas9 system and three phenotype-based screening strategies. We have utilized the CENP-E knockout cell line to establish a novel approach to validate the specificity and toxicity of the CENP-E inhibitors, which is useful for drug development and biological research. Biology Functional Assessment of Kinesin-7 CENP-E in Spermatocytes Using In Vivo Inhibition, Immunofluorescence and Flow Cytometry Meng-Fei Xu1,2, Yu-Hao Yang1,2, Ya-Lan Wei3,4, Jing-Lian Zhang1,2, Xuan Lin1,2, Xiao-Yan Lin1,2, Hui Chen1,2, Zhen-Yu She1,2 1Department of Cell Biology and Genetics, The School of Basic Medical Sciences, Fujian Medical University, 2Key Laboratory of Stem Cell Engineering and Regenerative Medicine, Fujian Province University, 3Fujian Obstetrics and Gynecology Hospital, 4Medical Research Center, Fujian Maternity and Child Health Hospital, Affiliated Hospital of Fujian Medical University This article reports an in vivo inhibition of CENP-E through abdominal surgery and testicular injection of GSK923295, a valuable model for male meiotic division. Using the immunofluorescence, flow cytometry and transmission electron microscopy assays, we show that CENP-E inhibition results in chromosome misalignment and genome instability in mouse spermatocytes.
