Centromere-Associated Protein-E Gene Editing Using the CRISPR/Cas9 System

June 23, 2023

使用 CRISPR/Cas9 系统生成着丝粒相关蛋白- E ^CENP-E-/- 敲除细胞系

Transfection, Isolation, and Screening of the CENP-E Knockout HeLa Cells

Immunofluorescence Staining and High-Resolution Confocal Microscopy of CRISPR/Cas9-Mediated CENP-E Knockout HeLa Cells

Chromosome Preparation and Karyotype Analysis of CRISPR/Cas9-Mediated CENP-E Knockout HeLa Cells

Meng-Fei Xu², Jie Chen², Yue Xu², Jing-Lian Zhang², Yi Zhou², Jie-Jie He², Shan Wu², Ya-Lan Wei⁴, Zhen-Yu She²

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

Summary

本文报道了使用 CRISPR/Cas9 系统和三种基于表型的筛选策略构建着丝粒相关蛋白-E （CENP-E）敲除细胞的过程。我们利用 CENP-E基因敲除细胞系建立了一种新的方法来验证CENP-E抑制剂的特异性和毒性，这对药物开发和生物学研究很有用。

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