使用CRISPR/Cas9系统生成与中心粒相关蛋白-E CENP-E-/-淘汰细胞系
Meng-Fei Xu1, Jie Chen1, Yue Xu1
1Department of Cell Biology and Genetics, The School of Basic Medical Sciences, Fujian Medical University; Key Laboratory of Stem Cell Engineering and Regenerative Medicine, Fujian Province University.
Journal of visualized experiments : JoVE
|July 10, 2023
概括
研究人员使用CRISPR/Cas9基因编辑创建CENP-E淘汰HeLa细胞. 这种CENP-E淘汰模型揭示了线粒体缺陷,并有助于开发新的癌症药物.
科学领域:
- 细胞生物学 细胞生物学
- 遗传学 是一个遗传学.
- 分子生物学分子生物学
背景情况:
- 中心分子相关蛋白-E (CENP-E) 对于染色体对齐和组装检查点至关重要.
- 研究CENP-E的直接功能是具有挑战性的,因为细胞循环在被切除后会停止.
研究的目的:
- 通过使用CRISPR/Cas9.9,生成一个CENP-E淘汰 (CENP-E-/-) 人类HeLa细胞系.
- 建立有效的选方法来识别淘汰细胞.
- 研究CENP-E删除对线粒分裂的影响,并开发CENP-E抑制剂查模型.
主要方法:
- 使用CRISPR/Cas9基因编辑来创建CENP-E-/- HeLa细胞.
- 基于表型的查策略,包括殖民地查和染色体对齐和蛋白质光的分析,得到了优化.
- 已建立的细胞系被用来开发和验证识别CENP-E抑制剂的方法.
主要成果:
- 成功生成了CENP-E淘汰HeLa细胞.
- 优化的查方法提高了淘汰细胞识别的效率和成功率.
- 删除CENP-E导致染色体错位,BubR1局部异常和线粒异常.
- 使用淘汰赛模型建立了一种用于识别和验证CENP-E抑制剂的新方法.
结论:
- 克里斯普尔/卡斯9系统提供了一种强大的方法来产生CENP-E淘汰细胞.
- CENP-E-/- HeLa细胞系是研究细胞分裂机制的宝贵工具.
- 这种淘汰赛模型有助于发现和验证CENP-E抑制剂,以开发潜在的抗瘤药物.
更多相关视频
11:35Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells
Published on: June 16, 2017
12.6K
11:49Author Spotlight: Establishing CENP-E Knockout HeLa Cells – A Novel Approach to Study Kinesin-7 CENP-E Biology and its Inhibitors
Published on: June 23, 2023
754
相关概念视频
01:59CRISPR
Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced Short...
01:28CRISPR/Cas9 Genome Editing
The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...