Helq对干细胞多能性影响的研究

Exploration of the effect of Helq gene on stem cell pluripotency

目的探讨Helq缺失是否影响干细胞的多能性。方法利用CRISPR-Cas9基因编辑技术获得Helq敲除的胚胎干细胞。结果免疫荧光染色结果显示敲除Helq后,Oct4及Nanog的表达与对照组相比无明显差异,通过四倍体补偿实验证明胚胎干细胞的多能性不受影响。同时我们利用Helq敲除的胚胎干细胞继续进行体外分化,最终可以得到Day2的上胚层样细胞。通过免疫染色及real-time PCR分析,结果表明Helq敲除的胚胎干细胞分化为上胚层细胞液无明显异常。结论 Helq缺失不影响多能性干细胞的多能性。

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.