Intracellular selection of trans-cleaving hammerhead ribozymes

doi:10.1093/nar/gkz018

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	Intracellular selection of trans-cleaving hammerhead ribozymes
	Huang, Xin 2; Zhao, Yongyun 1; Pu, Qinlin 2; Liu, Getong 2; Peng, Yan 2; Wang, Fei 2; Chen, Gangyi 2; Sun, Meiling 2; Du, Feng 2; Dong, Juan 2
刊名	NUCLEIC ACIDS RESEARCH
	2019
卷号	47 期号:5 页码:2514-2522
ISSN号	0305-1048
DOI	10.1093/nar/gkz018
产权排序	1
文献子类	Article
英文摘要	Hammerhead ribozyme is the smallest and best characterized catalytic RNA-cleaving ribozyme. It has been reported as potential therapeutic tools to manipulate the expression of target genes. However, most of naturally occurring hammerhead ribozymes process self-cleavage rather than cleave substrate RNA in trans, and its high intracellular activity relies on the tertiary interaction of Loop II and steam I bulge, resulting in decreased performance as applied in gene silencing. We described a direct intracellular selection method to evolve hammerhead variants based on trans-cleavage mode via using a toxin gene as the reporter. And a dual fluorescence proteins system has also been established to quantitatively evaluate the efficiency of selected ribozymes in the cell. Based on this selection strategy, we obtained three mutants with enhanced intracellular cleaving activity compared to wide type hammerhead ribozyme. The best one, TX-2 was revealed to possess better and consistent gene knockdown ability at different positions on diverse targeted mRNA either in prokaryotic or eukaryotic cells than wild-type hammerhead ribozyme. These observations imply the efficiency of the intracellular selection method of the trans-acting ribozyme and the potentials of improved ribozyme variants for research and therapeutic purposes.
学科主题	Biochemistry & Biophysics
URL标识	查看原文
WOS关键词	CLEAVAGE ; DISCOVERY ; HIV ; RNA ; VARIANTS ; CELLS ; TRIAL
WOS研究方向	Biochemistry & Molecular Biology
语种	英语
出版者	OXFORD UNIV PRESS
WOS记录号	WOS:000467963700034
内容类型	期刊论文
源URL	[http://210.75.237.14/handle/351003/31067]
专题	国家天然药物工程技术研究中心_天然产物研究
作者单位	1.Sichuan Univ, West China Hosp, Lab Stem Cell Biol, State Key Lab Biotherapy, Chengdu 610041, Sichuan, Peoples R China 2.Chinese Acad Sci, Nat Prod Res Ctr, Chengdu Inst Biol, Chengdu 610041, Sichuan, Peoples R China;
推荐引用方式 GB/T 7714	Huang, Xin,Zhao, Yongyun,Pu, Qinlin,et al. Intracellular selection of trans-cleaving hammerhead ribozymes[J]. NUCLEIC ACIDS RESEARCH,2019,47(5):2514-2522.
APA	Huang, Xin.,Zhao, Yongyun.,Pu, Qinlin.,Liu, Getong.,Peng, Yan.,...&Mo, Xianming.(2019).Intracellular selection of trans-cleaving hammerhead ribozymes.NUCLEIC ACIDS RESEARCH,47(5),2514-2522.
MLA	Huang, Xin,et al."Intracellular selection of trans-cleaving hammerhead ribozymes".NUCLEIC ACIDS RESEARCH 47.5(2019):2514-2522.