髓细胞条件性METTL3基因敲除小鼠的构建与鉴定
Journal of Zhengzhou University(Medical Sciences)（2022）
Abstract
目的:构建髓细胞条件性METTL3基因敲除小鼠模型.方法:将Lyz2-cre(+/+)雌性小鼠与MET-TL3(flox/flox)雄性小鼠杂交,得到METTL3(flox/-)Lyz2-cre(+/-)杂合子小鼠,该杂合子小鼠自交,得到METTL3(flox/flox)Lyz2-cre(+/+)小鼠.提取小鼠鼠尾组织DNA,经PCR扩增、琼脂糖凝胶电泳和DNA序列测定,在DNA水平判断小鼠基因型;提取小鼠骨髓细胞RNA,利用RT-PCR法检验METTL3的敲除效率;利用成像流式细胞仪在蛋白水平分析髓细胞中METTL3的敲除效率.结果 与结论:在DNA、RNA和蛋白水平分别验证了髓细胞条件性METTL3基因敲除小鼠模型构建成功,该小鼠存活并且可育,为研究METTL3在髓源抑制性细胞功能中的作用提供了研究平台.
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