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

June 23, 2023

Generering af centromer-associeret protein-E CENP-E^{-/- knockout-cellelinjer} ved hjælp af CRISPR/Cas9-systemet

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

Denne artikel rapporterer konstruktionen af centromer-associeret protein-E (CENP-E) knockout-celler ved hjælp af CRISPR / Cas9-systemet og tre fænotypebaserede screeningsstrategier. Vi har brugt CENP-E knockout-cellelinjen til at etablere en ny tilgang til validering af CENP-E-hæmmernes specificitet og toksicitet, hvilket er nyttigt til lægemiddeludvikling og biologisk forskning.