Research on the knockout of LMNA gene by CRISPR/Cas9 system in human cell lines
Research on the knockout of LMNA gene by CRISPR/Cas9 system in human cell lines
复制标题
DOI：
10.16288/j.yczz.18-146
复制
发表时间：
2019-01-01
期刊：
影响因子：
--
通讯作者：
Shu, Wei
中科院分区：
文献类型：
--
作者：
Liu, Heng;Li, Dongming;Shu, Wei
LMNA基因编码核核纤层蛋白A和核纤层蛋白C，与核膜组织、基因组稳定性和细胞分化有关。LMNA的异常表达普遍存在于人类肿瘤中，并且其突变导致各种形式的核纤层蛋白病，包括Emery-Dreifuss肌营养不良症（EDMD）、扩张型心肌病（DCM）和Hutchinson-Gliford早衰综合征（HGPS）。为了进一步确定LMNA基因在细胞生理学中的功能，本研究使用CRISPR/Cas9技术体外编辑293 T和HepG 2细胞的LMNA基因，从而产生两种稳定的LMNA基因敲除（LMNA KO）细胞系。与相应的野生型细胞相比，LMNA KO细胞系显示增殖能力降低、凋亡增加、细胞形态改变和核膜结构不均匀。本研究首次报道了LMNA KO永生化细胞系的构建及其形态学变化，为进一步研究LMNA基因功能和致病性突变奠定了基础。
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.