中国组织工程研究 ›› 2016, Vol. 20 ›› Issue (18): 2725-2730.doi: 10.3969/j.issn.2095-4344.2016.18.021

• 动物模型综述 Animal models review • 上一篇    下一篇

时间特异性基因敲除:技术、造模及现状和未来

陈瑞君,黄晓峰,张方明   

  1. 首都医科大学附属北京友谊医院口腔科,北京市 100050
  • 收稿日期:2016-02-16 出版日期:2016-04-29 发布日期:2016-04-29
  • 通讯作者: 黄晓峰,博士,教授,主任医师,首都医科大学附属北京友谊医院口腔科,北京市 100050
  • 作者简介:陈瑞君,女,1986年生,甘肃省兰州市人,汉族,首都医科大学附属北京友谊医院口腔科在读硕士研究生,主要从事口腔发育生物学研究。
  • 基金资助:

    国家自然科学基金项目(81170943),北京市自然科学基金项目(7162053)

Time-specific gene knockout technology: model establishment, present and future 

Chen Rui-jun, Huang Xiao-feng, Zhang Fang-ming   

  1. Department of Stomatology, Beijing Friendship Hospital, Capital Medical University, Beijing 100050, China
  • Received:2016-02-16 Online:2016-04-29 Published:2016-04-29
  • Contact: Huang Xiao-feng, D.D.S., Ph.D., Professor, Chief physician, Department of Stomatology, Beijing Friendship Hospital, Capital Medical University, Beijing 100050, China
  • About author:Chen Rui-jun, Studying for master’s degree, Department of Stomatology, Beijing Friendship Hospital, Capital Medical University, Beijing 100050, China
  • Supported by:

    the National Natural Science Foundation of China, No. 81170943; the Natural Science Foundation of Beijing City, No. 7162053

摘要:

文章快速阅读:

 

文题释义:
Cre/loxp系统:包括Cre重组酶和Loxp位点两个部分,Cre重组酶由大肠杆菌噬菌体P1的Cre基因编码,由343个氨基酸组成,不仅具有催化活性,而且跟限制酶相似,识别特异的DNA序列,如loxp位点,引起重组。
基因敲除:是20世纪80年代发展起来的,是建立在基因同源重组技术基础以及胚胎干细胞技术基础上的一种新分子生物学技术。基因敲除通过同源重组将外源基因定点整合入靶细胞基因组上某一确定的位点,以达到定点修饰改造染色体上某一基因的目的。基因敲除克服了随机整合的盲目性和偶然性,是一种理想的修饰、改造生物遗传物质的方法。基因敲除技术主要应用于动物模型的建立,而最成熟的实验动物是小鼠,近年来,牛、羊、猪、猴等大型哺乳动物实现了基因敲除。
 
背景:传统基因敲除技术不能在特定时间敲除目标基因,动物模型往往在胚胎发育时期或出生后死亡,限制了基因功能的研究。
目的:综述时间特异性基因敲除技术的研究现状。
方法:以“gene knockout,inducer,Cre/loxP system”为关键词检索建库至2014年8月PubMed数据库。从初检文献中排除重复实验和与研究目的无关的文献,通过分析结果对时间特异性基因敲除技术的研究做出总结。
结果与结论:大多数个体全部细胞基因敲除动物在出生后往往不能存活,它们因为某个重要基因缺失后胚胎早期产生严重生理性缺陷而死亡。因此,对于某个基因在特定时间的功能、生物个体在特定发育时间段内的遗传特征、以及某些获得性疾病在特定时间内的治疗等一系列问题,由于研究方法的局限性至今进展有限。而条件性基因敲除动物,尤其是时间特异性基因敲除动物的出现为这些领域的研究提供了研究模型。文章总结了近几年国内外关于时间特异性基因敲除动物的研究、发展及其应用,阐明特定基因的时间特异性基因敲除技术在科学研究领域的重大意义和广阔前景。

中国组织工程研究杂志出版内容重点:肾移植;肝移植;移植;心脏移植;组织移植;皮肤移植;皮瓣移植;血管移植;器官移植;组织工程
ORCID: 0000-0002-6850-0250(黄晓峰)

关键词: 实验动物, 基因敲除, 遗传, 同源重组, 位点特异性, Cre/loxP系统, 诱导剂, 动物模型, 综述

Abstract:

BACKGROUND: The target gene cannot be deleted at specific time by using conditional gene knockout system. Most gene knockout animals cannot survive during embryonic development and after birth, which limits the study of gene function.

OBJECTIVE: To review the recent advances about time-specific gene knockout technology.
METHODS: An online search in PubMed was performed from the initial stage of database establishment to August 2014. The key words were gene knockout, inducer and Cre/loxP system. The repeated experiment and unrelated articles were excluded during primary search. A summary was made by analyzing time-specific gene knockout technology.
RESULTS AND CONCLUTION: Most important gene knockout animals could not survive after birth, or die at early embryonic stage due to serious physical defects induced by an important gene deletion. Therefore, the progress in the functionality of a gene at a particular time, the genetic characteristics of an individual organism in a particular developmental period, and the treatment of certain acquired disease at a given time have been limited because of the limitations of research methods. However, the occurrence of conditional gene knockout animals, especially time-specific knockout animals, provides a research model for the study of these areas. This study reviewed the recent advances, development and application about time-specific gene knockout animals, and discussed the significance and broad prospects of time-specific gene knockout technology of specific genes in the field of scientific research. 
中国组织工程研究杂志出版内容重点:肾移植;肝移植;移植;心脏移植;组织移植;皮肤移植;皮瓣移植;血管移植;器官移植;组织工程

Key words: Gene Knockout Techniques, Gene Deletion, Chimera, Cell Division