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Hereditas(Beijing) >

2019 , Vol. 41 >Issue 1: 66 - 75

DOI: https://doi.org/10.16288/j.yczz.18-146

Research on the knockout of LMNA gene by CRISPR/Cas9 system in human cell lines

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  • 1. Department of Cell Biology and Genetics, Basic Medical College, Guangxi Medical University, Nanning 530021, China
    2. College of Environmental Resources, Guangxi Normal University, Guilin 541004, China

Received date: 2018-09-13

  Revised date: 2018-11-21

  Online published: 2018-12-06

Supported by

[Supported by the National Natural Science Foundation of China (No. 31660311) and Guangxi Medical University Innovation and Entrepreneurship Project (No. 201710598055)]

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Abstract

The LMNA gene encodes the nuclear Lamin A and Lamin C proteins, and is related to nuclear membrane organization, genome stability and cell differentiation. Abnormal expression of LMNA is ubiquitous in human tumors, and its mutation leads to various forms of laminopathies, including Emery-Dreifuss muscular dystrophy (EDMD), dilated cardiomyopathy (DCM), and Hutchinson-Gliford progeria syndrome (HGPS). To further determine the functions of the LMNA gene in cellular physiology, the present study used the CRISPR/Cas9 technique to edit the LMNA gene of 293T and HepG2 cells in vitro, which resulted in two stable LMNA gene knockout (LMNA KO) cell lines. Compared to the respective wild type cells, the LMNA KO cell lines showed decrease in proliferation ability, increase in apoptosis, alteration in cellular morphology and uneven structures in the nucleus membrane. In this study, we report for the first time the results on the construction of LMNA KO immortalized cell lines and characterization of their morphological changes, thereby laying the foundation for the further studies of the LMNA gene functions and pathogenic mutations.

Key words： LMNA gene; CRISPR/Cas9; 293T; HepG2; cell morphology

Cite this article

Heng Liu, Dongming Li, Lanyu Zhu, Lejin Lai, Wanyun Yan, Yushuang Lu, Yi Wei, Yueqi Huang, Mei Fang, Yuangang Su, Fang Yang, Wei Shu . Research on the knockout of LMNA gene by CRISPR/Cas9 system in human cell lines[J]. Hereditas(Beijing), 2019 , 41(1) : 66 -75 . DOI: 10.16288/j.yczz.18-146

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