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戊二酸尿症Ⅰ型基因敲除大鼠模型的构建和鉴定 被引量：2

Establishment and identification of a glutaric aciduria typeⅠgene knockout rat model
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摘要 目的:构建戊二酰辅酶A脱氢酶(GCDH)基因敲除大鼠,为进一步研究戊二酸尿症Ⅰ型发病机制和治疗方法提供动物模型。方法:利用转录激活因子样效应物核酸酶(TALEN)技术制备GCDH基因敲除大鼠,设计GCDH基因打靶位点,构建载体,体外转录mRNA,显微注射入SD大鼠受精卵,鉴定F0代大鼠。经饲养、配种、繁殖后,剪尾、PCR及电泳检测子代大鼠基因型,观察并统计GCDH基因敲除幼鼠出生及存活情况。高赖氨酸饮食诱导GCDH-/-大鼠发病,HE染色观察大鼠脑组织的病理变化。结果:经基因检测及测序验证,成功制备GCDH-/-大鼠,子代大鼠基因型符合孟德尔遗传定律并可稳定繁殖。8周龄GCDH-/-大鼠给予高赖氨酸饲料喂养4周,5只中存活4只;同等条件的5只野生大鼠全部存活率。与野生型相比,高赖氨酸饮食GCDH-/-大鼠脑出现细胞水肿、排列紊乱和空泡样变性。结论:成功建立GCDH-/-大鼠模型。 Aim:To construct and identify a GCDH gene knockout rat,which could provide animal models to study the pathogenesis and treatments for glutaric aciduria typeⅠ.Methods:GCDH knockout rats were created by using transcription-activator-like effector nuclease(TALEN)technology.GCDH gene targeting sites were designed and then the vectors were constructed.mRNA was microinjected into SD rat zygotes.F0 rats were verified by DNA level.After feeding,mating,and breeding,the genotypes of the offsprings were analyzed by PCR and electrophoresis.The birth and survival of GCDH-/-pups were recorded.Brain pathological changes of GCDH-/-rats with high lysine diet were observed by HE staining.Results:After genetic testing and sequencing verification,GCDH-/-rats were successfully established,and the genotypes of the offspring rats conform to Mendelian laws of inheritance and could reproduce stably.Eight-week-old GCDH-/-rats were fed with high lysine diet for 4 weeks,and 4 out of 5 rats survived;5 wild type rats under the same conditions all survived.HE staining showed that the brain cells of GCDH-/-rats with high lysine diet had edema,disordered arrangement and vacuolar degeneration compared with that of wild type rats.Conclusion:The GCDH-/-rat model has been successfully established.
作者 田凤艳 李艳云 甄诚 顾朝辉 TIAN Fengyan;LI Yanyun;ZHEN Cheng;GU Chaohui(Department of Pediatrics,the First Affiliated Hospital,Zhengzhou University,Zhengzhou 450052;Academy of Medical Sciences,Zhengzhou University,Zhengzhou 450052;Department of Urinary Surgery,the First Affiliated Hospital,Zhengzhou University,Zhengzhou 450052)
机构地区 郑州大学第一附属医院儿科 郑州大学医学科学院 郑州大学第一附属医院泌尿外科
出处 《郑州大学学报（医学版）》 CAS 北大核心 2020年第6期801-805,共5页 Journal of Zhengzhou University(Medical Sciences)
基金 国家自然科学基金项目(81200883) 河南省医学科技攻关项目(201702015)。
关键词 戊二酸尿症Ⅰ型 转录激活因子样效应物核酸酶技术 基因敲除 戊二酰辅酶A脱氢酶基因 大鼠 glutaric aciduria typeⅠ transcription activator-like effector nuclease technology gene knockout GCDH gene rat
分类号 R725.8 [医药卫生—儿科]
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郑州大学学报（医学版）
2020年 第6期

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