p53基因敲除大鼠模型的构建及表型分析

doi:10.13560/j.cnki.biotech.bull.1985.2018-0086

生物技术通报 ›› 2018, Vol. 34 ›› Issue (8): 170-174.doi: 10.13560/j.cnki.biotech.bull.1985.2018-0086

p53基因敲除大鼠模型的构建及表型分析

霍桂桃¹, 杨艳伟¹, 吴曦², 刘甦苏², 李芊芊¹, 周舒雅², 柳全明², 王三龙¹, 沈月雷³, 吕建军¹, 范昌发²

1. 中国食品药品检定研究院国家药物安全评价监测中心药物非临床安全评价研究北京市重点实验室,北京 100176;
2. 中国食品药品检定研究院实验动物资源研究所模式动物研究室,北京 100050;
3. 百奥赛图基因生物技术有限公司,北京 101111

收稿日期:2018-01-23 出版日期:2018-08-26 发布日期:2018-09-04
作者简介:霍桂桃,女,博士,副主任药师,研究方向：临床前药物安全性评价毒性病理学诊断及分子病理学,E-mail：huoguitao@nifdc.org.cn;杨艳伟为共同第一作者
基金资助:
国家青年自然基金项目（81502396）

Construction and Phenotypic Analysis of Gene p53 Knockout Rat Model

HUO Gui-tao¹, YANG Yan-wei¹, WU Xi², LIU Su-su², LI Qian-qian¹, ZHOU Shu-ya², LIU Quan-ming², WANG San-long¹, SHEN Yue-lei³, LÜ Jian-jun¹, FAN Chang-fa²

1. Beijing Key Laboratory for Safety Evaluation of Drugs,National Center for Safety Evaluation of Drugs,National Institutes for Food and Drug Control,Beijing 100176;
2. Division of Animal Model Research,Institute for Laboratory Animal Resources,National Institutes for Food and Drug Control,Beijing 100050;
3. Beijing Biocytogen Co. Ltd.,Beijing 101111

Received:2018-01-23 Published:2018-08-26 Online:2018-09-04

摘要/Abstract

摘要： 使用TALEN技术制作p53基因敲除大鼠模型,建立种群并分析大鼠表型。设计特异性识别p53基因外显子2的TALEN蛋白并构建相应载体,将体外转录的TALEN mRNA注射SD大鼠受精卵,出生后从仔鼠中通过测序筛选靶位点正确剪切的阳性小鼠。结果显示,注射得到11只新生仔鼠,通过测序发现其中有10只靶位点发生剪切。选取其中4只作为首建鼠进行繁殖。p53基因敲除纯合子表现出肿瘤易发性,主要自发性肿瘤类型为恶性纤维组织肉瘤。此外,p53基因敲除纯合子大鼠表现出眼睛发育异常,视网膜变性及晶状体纤维排列紊乱。通过TALEN技术高效地获得了p53基因敲除大鼠模型,纯合子敲除大鼠除了易发肿瘤并伴有眼发育异常,因而该敲除大鼠模型除了可用于研究肿瘤发生机制外,也可用于研究p53基因在眼发育过程中的功能。

关键词: TALEN, p53基因敲除, SD大鼠, 肿瘤, 眼发育异常

Abstract: The objectives of this work are to construct rat model with gene p53 knockout using TALEN,to establish colony,and to analyze the phenotypes of the rats. TALEN protein specifically recognizing exon 2 of gene p53 was designed and the corresponding vectors were constructed. In vitro transcribed TALEN mRNA was injected into fertilized eggs of SD rats. New born rats with right splicing at the target site were determined by sequencing. Together 11new-born rats were obtained after injection and 10 of them were having right splicing at target site by sequencing. Four rats were selected as founder to establish the colony. The homozygotes with gene p53 knockout presented tumor susceptibility and the main type of spontaneous tumor was malignant fibrous histiocytoma. In addition,the homozygous rats with gene p53 knockout also had eye developmental defects,such as retina degeneration and disorganized fiber of lens. In conclusion,rat model with p53knockout is efficiently constructed by TALEN,the model exhibits tumor susceptibility and eye developmental defects,indicating that the model can be used for studying the mechanisms of tumorigenesis and the function of gene p53 in eye development.

Key words: TALEN, gene p53 knockout, SD rat, tumor, eye developmental defect

霍桂桃, 杨艳伟, 吴曦, 刘甦苏, 李芊芊, 周舒雅, 柳全明, 王三龙, 沈月雷, 吕建军, 范昌发. p53基因敲除大鼠模型的构建及表型分析[J]. 生物技术通报, 2018, 34(8): 170-174.

HUO Gui-tao, YANG Yan-wei, WU Xi, LIU Su-su, LI Qian-qian, ZHOU Shu-ya, LIU Quan-ming, WANG San-long, SHEN Yue-lei, LÜ Jian-jun, FAN Chang-fa. Construction and Phenotypic Analysis of Gene p53 Knockout Rat Model[J]. Biotechnology Bulletin, 2018, 34(8): 170-174.

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p53基因敲除大鼠模型的构建及表型分析

Construction and Phenotypic Analysis of Gene p53 Knockout Rat Model

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