利用CRISPR-dCas9系统促进脂肪干细胞的成骨诱导分化  

Control of osteogenic differentiation in adipose-derived stem cells via endogenous gene activation using CRISPR-dCas9

作　　者：杨馥如 王学智[3] 余四九[1] YANG Fu-ru;WANG Xue-zhi;YU Si-jiu(College of Veterinary Medicine,Gansu Agricultural University,Lanzhou 730050,China;Shanghai Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Shanghai 200240,China;Lanzhou Institute of Husbandryand Pharmaceutical Sciences,Chinese Academy of Agricultural Sciences,Lanzhou 730050,China)

机构地区：[1]甘肃农业大学动物医学院,甘肃兰州730050 [2]中国农业科学院上海兽医研究所,上海200241 [3]中国农业科学院兰州畜牧与兽药研究所,甘肃兰州730050

出　　处：《中国兽医科学》2019年第3期372-377,共6页Chinese Veterinary Science

基　　金：国家科技基础性工作专项(2013FY110600-6)

摘　　要：为了建立一种快速促进脂肪间充质干细胞成骨诱导分化的方法,本研究利用CRISPR-dCas9系统快速激活成骨前体基因的表达,通过对细胞内前体基因转录的检测和诱导分化后茜素红染色来确定成骨诱导分化的效率。结果可见,通过CRISPR-dCas9系统可以快速地启动RUNX2和OSX基因的转录;经成骨诱导分化后茜素红染色显示,基因编辑过的细胞成骨诱导更快。本研究结果表明,通过CRISPR-On系统可以有效地促进成骨前体细胞相关基因RUNX2和OSX的表达,从而使得骨前体细胞相关基因可以被快速地内源性激活,使得脂肪间充质干细胞快速进入成骨诱导分化状态,促进成骨分化的潜能。To establish a method of osteogenic differentiation rapidly,CRISPR-d Cas9 was used to endogenously promote the transcription of osteogenic progenitor genes in adipose-derived stem cell.The cells were identified by transcription levels and morphology analysis by RT-q PCR and Alzarin red staining.In result,osteogenic progenitor genes could be upregulated,then the cells could differentiate towards the osteogenic lineage rapidly,indicating endogenous activation of osteogenic progenitor genes(RUNX2 and OSX)through the CRISPR-on system.The above-mentioned results showed that osteogenic progenitor genes can be effectively activated,which promote the ability of osteogenic differentiation.

关 键 词：脂肪干细胞 成骨诱导分化 CRISPR-On 

分 类 号：S852.161[农业科学—基础兽医学]