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作 者:崔古贞 花登雄 谷俊莹 陈峥宏 CUI Guzhen;HUA Dengxiong;GU Junying;CHEN Zhenghong(Department of Microbiology,School of Basic Medicine,Guizhou Medical University,Guiyang 550025,Guizhou,China;School of Clinical Laboratory Science,Guizhou Medical University,Guiyang 550004,Guizhou,China;Key Laboratory of Microbiology and Parasitology of Education Department of Guizhou,School of Basic Medicine,Guizhou Medical University,Guiyang 550025,Guizhou,China)
机构地区:[1]贵州医科大学基础医学院微生物学教研室,贵州贵阳550025 [2]贵州医科大学医学检验学院,贵州贵阳550004 [3]贵州医科大学基础医学院病原生物学重点实验室,贵州贵阳550025
出 处:《生物工程学报》2022年第3期915-924,共10页Chinese Journal of Biotechnology
基 金:国家自然科学基金(31760318);贵州省科技计划项目([2019]1441,[2018]5779-17)。
摘 要:型内含子(groupⅡintrons)是一类具有自我剪接功能的核酶,能够通过“归巢”(retrohoming)机制高频插入到DNA靶位点。Ⅱ型内含子对DNA靶位点的识别和剪接具有高度专一性和高效性,这种特性使其在基因工程中具有重要的应用价值。文中首先综述了Ⅱ型内含子基因打靶原理及其在微生物遗传改造中的应用;然后,根据Ⅱ型内含子“归巢”特点及其依赖高浓度Mg2+的特性,分析了Ⅱ型内含子在多功能基因编辑及真核生物应用中的局限性;最后,以笔者课题组研究工作为基础,结合Ⅱ型内含子自身结构特点,分析了Ⅱ型内含子在新型基因编辑工具开发方面的潜能,为Ⅱ型内含子生物技术应用提供借鉴。GroupⅡintrons are self-splicing ribozymes,which insert directly into target sites in DNA with high frequency through“retrohoming”.They specifically and efficiently recognize and splice DNA target sites,endowing themselves with great potential in genetic engineering.This paper reviewed the gene targeting principle of groupⅡintrons and the application in microbial genetic modification,and then analyzed the limitations of them in multi-functional gene editing and eukaryotes based on the“retrohoming”characteristics and the dependence on high Mg2+concentration.Finally,we dissected the potential of groupⅡintrons in the development of novel gene editing tools based on our previous research outcome and the structural characteristics of the introns,hoping to provide a reference for the application of groupⅡi ntrons in biotechnology.
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