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

June 23, 2023

Geração de Linhagens Celulares Knockout da Proteína Associada ao Centrômero-E ^CENP-E-/- utilizando o Sistema CRISPR/Cas9

02:19 min

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

04:10 min

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

03:07 min

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

02:19 min

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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

Este artigo relata a construção de células knockout da proteína E associada a centrômeros (CENP-E) usando o sistema CRISPR/Cas9 e três estratégias de triagem baseadas em fenótipos. Utilizamos a linhagem celular knockout do CENP-E para estabelecer uma nova abordagem para validar a especificidade e toxicidade dos inibidores do CENP-E, o que é útil para o desenvolvimento de fármacos e pesquisa biológica.