Oturum Aç

Fujian Maternity and Child Health Hospital

2 ARTICLES PUBLISHED IN JoVE

Biology

Generation of Centromere-Associated Protein-E CENP-E^-/- Knockout Cell Lines using the CRISPR/Cas9 System

Meng-Fei Xu^1,2, Jie Chen^1,2, Yue Xu^1,2, Jing-Lian Zhang^1,2, Yi Zhou^1,2, Jie-Jie He^1,2, Shan Wu^1,2, Ya-Lan Wei^3,4, Zhen-Yu She^1,2

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

Epon Post Embedding Correlative Light and Electron Microscopy

Shuyuan Wang^*1, Haiyan Xiong^*1,2, Qiyuan Chang^*1,3, Xudong Zhuang^*4,5,6, Yaochen Wu^1,3, Xinrui Wang^4,5,6, Congxian Wu¹, Zhifei Fu^1,7,8

¹Public Technology Service Center, Fujian Medical University, ²The School of Pharmacy, Fujian Medical University, ³The School of Basic Medical Sciences, Fujian Medical University, ⁴College of Clinical Medicine for Obstetrics & Gynecology and Pediatrics, Fujian Medical University, ⁵Medical Research Center, Fujian Maternity and Child Health Hospital, ⁶NHC Key Laboratory of Technical Evaluation of Fertility Regulation for Non-human Primate, Fujian Maternity and Child Health Hospital, ⁷Institute of Neuroscience, Fujian Medical University, ⁸Fujian Key Laboratory of Molecular Neurology, Fujian Medical University

We present a detailed protocol for Epon post-embedding correlative light and electron microscopy using a fluorescent protein called mScarlet. This method can maintain the fluorescence and the ultrastructure simultaneously. This technique is amenable to a wide variety of biological applications.