去泛素化酶OTUD6A基因敲除小鼠的构建及表型初步分析

Generation and phenotype analysis of knockout mice of deubiquitinase OTUD6A

目的建立去泛素化酶OTUD6A基因敲除小鼠模型,并对该模型进行初步表型分析。方法基于CRISPR/Cas9基因编辑技术构建OTUD6A基因敲除小鼠;利用PCR、qPCR和Western blot技术(WB)分别在DNA水平、mRNA水平和蛋白水平上检测OTUD6A基因的表达;统计雄性OTUD6A纯合敲除(OTUD6A-/-)小鼠和雌性OTUD6A杂合子(OTUD6A+/-)小鼠交配所得后代各基因型小鼠比例及生长发育情况,对小鼠各组织器官利用苏木素-伊红染色(HE染色)进行病理学分析。结果成功建立OTUD6A基因敲除小鼠模型,该基因敲除小鼠可正常生长繁殖并符合伴性遗传定律。与同窝野生型(WT)小鼠相比,纯合敲除(OTUD6A-/-)小鼠在体型和体质量上无显著差异,主要组织脏器也无明显的病理学异常。结论成功构建OTUD6A基因敲除小鼠模型。纯合敲除(OTUD6A-/-)小鼠可正常生长发育,与WT小鼠相比无显著差异。

Objective To establish a mouse model of OTUD6 A gene knockout and to analyze the phenotype of the mouse model.Methods OTUD6 A gene knockout mice were constructed with CRISPR/Cas9 gene editing technology.PCR was used to detect the genotype of OTUD6 A knockout and wild type mice.The levels of OTUD6 A mRNA and protein were detected by qPCR and Western blot(WB),respectively.Male OTUD6 A homozygous knockout(OTUD6 A-/-)mice mated with female OTUD6 A heterozygous(OTUD6 A+/-)mice before the proportion,growth and development of the offspring were observed.Hematoxylin-eosin(H&E)staining was used to analyze the pathological change in all tissues and organs of the offspring.Results An OTUD6 A gene knockout mouse model was successfully established,and the mice could grow and reproduce normally in accordance with the law of sex inheritance.Compared with wild-type mice in the same litter,OTUD6 A homozygous knockout(OTUD6 A-/-)mice showed no significant difference in body size or relative body mass,and there were no obvious pathological abnormalities in the main tissues and organs.Conclusion An OTUD6 A gene knockout mouse model is successfully generated.OTUD6 A homozygous knockout(OTUD6 A-/-)mice show normal growth and development,which is not significantly different from wild-type mice.