Exploration of the effect of Helq gene on stem cell pluripotency
Received:January 13, 2016  
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DOI:10.3969/j.issn.1005-4847.2016.04.005
KeyWord:Helq;Pluripotency;Embryonic stem cells;Epiblast-like cells;CRISPR-Cas9
              
AuthorInstitution
万聪 教育部蛋白质化学和鱼类发育生物学重点实验室, 湖南师范大学生命科学学院, 长沙 ;发育生物学教研室, 南方医科大学基础医学院, 广州
黄亚萍 教育部蛋白质化学和鱼类发育生物学重点实验室, 湖南师范大学生命科学学院, 长沙 ;发育生物学教研室, 南方医科大学基础医学院, 广州
汪妹 教育部蛋白质化学和鱼类发育生物学重点实验室, 湖南师范大学生命科学学院, 长沙 ;发育生物学教研室, 南方医科大学基础医学院, 广州
肖亚梅 教育部蛋白质化学和鱼类发育生物学重点实验室, 湖南师范大学生命科学学院, 长沙
赵小阳 发育生物学教研室, 南方医科大学基础医学院, 广州
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Abstract:
      Objective To explore whether Helq deletion affect the pluripotency of stem cells. Methods Helq knockout embryonic stem cells were obtained by CRISPR-Cas9 gene editing technique.Results The results of immunofluorescence analysis showed that the expression of Oct4 and Nanog had no obvious difference to that of the control cells. The Helq-/- embryonic stem cells could produce viable pups by tetraploid complementation, indicating that their pluripotency was not affected. Meanwhile, we found that day 2 epiblast-like cells also were obtained through differentiation of the Helq-/- embryonic stem cells in vitro. Immunostaining and real-time PCR analysis showed that the gene expression of Helq-/- epiblast cells were similar to the wild type cells. Conclusions Taken together, it is proved that the genomic instability caused by Helq deletion does not affect the pluripotency of pluripotent stem cells.
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