期刊文献+
共找到10篇文章
< 1 >
每页显示 20 50 100
Application of Artificially Induced Double-strand Breaks (DSB) and Triplex-forming Oligonucleotides (TFO) in the Improvement of Gene Targeting Efficiency
1
作者 Hegang LI Wenke CHENG +5 位作者 Ke JIANG Xiaoli REN Yongping JIANG Lele HOU Xiaojing HAO Jinshan ZHAO 《Agricultural Biotechnology》 CAS 2013年第1期1-6,12,共7页
Gene targeting technology is an important means to investigate gene functions, but its efficiency of gene targeting is very low, especially for somatic cell targeting. Artificially induced double-strand breaks (DSB)... Gene targeting technology is an important means to investigate gene functions, but its efficiency of gene targeting is very low, especially for somatic cell targeting. Artificially induced double-strand breaks (DSB) and triplex forming oligonucleotide (TFO) are currently developed methods to improve the targeting efficiency. This paper summarized the basic principles, design ideas and application in gene targeting efficiency improvement of these two methods, analyzed and com- pared their characteristics, and finally proposed prospects for their future development. 展开更多
关键词 Gene targeting double-strand breaks Zinc finger nuclease Homing endonuclease Triplex-forming oligonucleotides
下载PDF
Blocking Translation of Oncogenic mRNA
2
作者 Kelvin N. Christie 《Journal of Cancer Therapy》 2023年第6期233-256,共24页
Double-stranded RNA-mediated interference (RNAi), antisense oligonucleotides (ASO), and ribozymes have excellent specificity to their target oncogenic mRNA. They also seem to show great promise when it comes to treati... Double-stranded RNA-mediated interference (RNAi), antisense oligonucleotides (ASO), and ribozymes have excellent specificity to their target oncogenic mRNA. They also seem to show great promise when it comes to treating cancer. The problem is that RNAi, ASO, and ribozymes have poor stability and are constantly being degraded by nucleases. Researchers have made some efforts to increase antisense oligonucleotides’ stability by creating phospharimidate and Phosphorothioate. Currently, ribozymes, antisense oligonucleotides, and (RNAi) are the three main methods used to target RNA. These methods are currently undergoing clinical trials for the purpose of focusing on specific RNAs involved in disorders like cancer and neurodegeneration. In fact, ASOs that target amyotrophic lateral sclerosis and spinal muscular atrophy have produced promising results in clinical trials. The formation of chemical alterations that boost affinity and selectivity while reducing noxiousness owing to off-target impacts are two benefits of ASOs. Another benefit is increased affinity. With a focus on RNAi and ASOs, this review illustrated the main therapeutic strategies of RNA therapy now in use. 展开更多
关键词 Antisense Oligonucleotides RIBOZYMES PHOSPHOROTHIOATE double-Stranded RNA-Mediated Interference nucleaseS
下载PDF
逆转录病毒载体介导的转基因鸡研究进展 被引量:3
3
作者 王令 张涛 尹亚军 《中国家禽》 北大核心 2015年第14期44-48,共5页
转基因鸡作为重要的经济动物和理想的生物反应器,具有广阔的应用前景。鸡基因组测序的完成加快了转基因鸡的研究和发展。病毒载体法是转基因鸡制备的主导方法之一,具有效率高和遗传稳定性良好的优势。本文对逆转录病毒载体特征、复制缺... 转基因鸡作为重要的经济动物和理想的生物反应器,具有广阔的应用前景。鸡基因组测序的完成加快了转基因鸡的研究和发展。病毒载体法是转基因鸡制备的主导方法之一,具有效率高和遗传稳定性良好的优势。本文对逆转录病毒载体特征、复制缺陷型病毒载体和复制整合双缺陷型病毒载体的优缺点及其介导的转基因鸡研究进行综述,展望人工核酸酶技术结合复制整合双缺陷型逆转录病毒载体在转基因鸡研究领域的应用前景。 展开更多
关键词 转基因鸡 逆转录病毒载体 复制整合双缺陷型病毒载体 人工核酸酶技术
下载PDF
CELⅠ酶对提高基因合成正确率的影响
4
作者 郝爱平 《湖北农业科学》 北大核心 2014年第22期5562-5564,共3页
利用CEL Ⅰ核酸内切酶能切割掉异源双链DNA分子中错配的单核苷酸碱基对的特性,研究了CELⅠ核酸内切酶是否能提高基因合成的正确率。结果表明,CEL Ⅰ核酸内切酶不仅能切割掉双链DNA分子中错配的单核苷酸碱基对,同时也能引入外源碱基对,... 利用CEL Ⅰ核酸内切酶能切割掉异源双链DNA分子中错配的单核苷酸碱基对的特性,研究了CELⅠ核酸内切酶是否能提高基因合成的正确率。结果表明,CEL Ⅰ核酸内切酶不仅能切割掉双链DNA分子中错配的单核苷酸碱基对,同时也能引入外源碱基对,使合成的目的双链DNA分子中错误率增加,不能达到提高基因合成正确率的目的。 展开更多
关键词 CELⅠ酶 双链DNA分子 基因合成正确率
下载PDF
供体同源臂长度对ZFN介导的同源重组效率的影响 被引量:6
5
作者 聂宇 乔艳乐 +1 位作者 陈瑶生 何祖勇 《中山大学学报(自然科学版)》 CAS CSCD 北大核心 2016年第4期100-107,共8页
应用锌指核酸酶(Zinc Finger Nuclease,ZFN)和序列同源的供体(Donor)作为模板,借助DNA的同源重组修复机制能够对动物基因组实现精确的遗传修饰。目前关于供体长度与ZFN介导的同源重组修复效率相关性的报道相对较少。本研究构建了一对靶... 应用锌指核酸酶(Zinc Finger Nuclease,ZFN)和序列同源的供体(Donor)作为模板,借助DNA的同源重组修复机制能够对动物基因组实现精确的遗传修饰。目前关于供体长度与ZFN介导的同源重组修复效率相关性的报道相对较少。本研究构建了一对靶向EGFP的ZFN并鉴定了活性,同时设计了一系列不同长度的供体,应用流式细胞分析术,在稳定整合了带有移码突变的EGFP基因的CHO细胞中系统分析了供体长度对ZFN介导的同源重组修复效率的影响。结果发现当同源臂单臂仅有50 bp时,即可有效支持ZFN介导的同源同组,随着同源臂长度的延伸,同源重组的效率有所提高,但要实现高效率的同源重组(较传统方法提高104倍),同源臂单臂长度需要延长至1 000 bp以上。这为今后如何设计合适的Donor,以提高ZFN等基因组编辑工具介导的同源重组效率提供了借鉴。 展开更多
关键词 锌指核酸酶 DNA双链断裂 同源重组 供体 同源臂
下载PDF
基于酶剪切量子点荧光放大技术的双元miRNA定量检测 被引量:1
6
作者 佟丽莹 洑颢 +4 位作者 贾志舰 梁照恒 祝远锋 甘棕松 周骏 《光子学报》 EI CAS CSCD 北大核心 2020年第5期137-146,共10页
将量子点荧光特性与双链特异性核酸酶的DNA剪切特性相结合,提出一种高灵敏度、高特异性的双元miRNA定量检测方案.首先,将量子点和四氧化三铁磁性纳米粒子分别与捕获DNA链接形成捕获探针,再与待测miRNA互补配对形成异源双链杂合结构,随... 将量子点荧光特性与双链特异性核酸酶的DNA剪切特性相结合,提出一种高灵敏度、高特异性的双元miRNA定量检测方案.首先,将量子点和四氧化三铁磁性纳米粒子分别与捕获DNA链接形成捕获探针,再与待测miRNA互补配对形成异源双链杂合结构,随后双链特异性核酸酶对杂合结构中的捕获DNA进行特异性剪切,实现量子点和待测miRNA从捕获探针分离,且分离的待测miRNA与捕获探针上未配对的DNA开始新一轮杂交和再剪切.经过上述循环过程,量子点从捕获探针大量释放,荧光信号不断增强,实现肿瘤标志物miRNA的高灵敏检测.实验结果表明,基于酶剪切量子点荧光放大技术,在1fmol/L至100pmol/L的浓度范围内,同时实现了肿瘤标志物miRNA-141及循环miRNA内参miRNA-1228的特异性定量检测,其检出限分别达到0.69fmol/L和0.21fmol/L.与实时荧光定量多聚核苷酸链式反应方法相比,该方案获得了相同的检测结果,且具有更高灵敏度. 展开更多
关键词 量子点 双链特异性核酸酶 MIRNA 荧光检测
下载PDF
Site-Specifc Gene Targeting Using Transcription Activator-Like Effector(TALE)-Based Nuclease in Brassica oleracea 被引量:3
7
作者 Zijian Sun Nianzu Li +6 位作者 Guodong Huang Junqiang Xu Yu Pan Zhimin Wang Qinglin Tang Ming Song Xiaojia Wang 《Journal of Integrative Plant Biology》 SCIE CAS CSCD 2013年第11期1092-1103,共12页
Site-specific recognition modules with DNA nuclease have tremendous potential as molecular tools for genome targeting. The type III transcription activator-like effectors (TALEs) contain a DNA binding domain consist... Site-specific recognition modules with DNA nuclease have tremendous potential as molecular tools for genome targeting. The type III transcription activator-like effectors (TALEs) contain a DNA binding domain consisting of tandem repeats that can be engineered to bind user-defined specific DNA sequences. We demonstrated that customized TALE-based nucleases (TALENs), constructed using a method called "unit assembly", specifically target the endogenous FRIGIDA gene in Brassica oleracea L. var. capitata L. The results indicate that the TALENs bound to the target site and cleaved double-strand DNA in vitro and in vivo, whereas the effector binding elements have a 23 bp spacer. The T7 endonuclease I assay and sequencing data show that TALENs made double-strand breaks, which were repaired by a non- homologous end-joining pathway within the target sequence. These data show the feasibility of applying customized TALENs to target and modify the genome with deletions in those organisms that are still in lacking gene target methods to provide germplasms in breeding improvement. 展开更多
关键词 Brassica oleracea double-strand break FRIGIDA gene targeting type III transcription activator-like effector-based nucleases non-homologous end-joining.
原文传递
Gene editing for corneal disease management
8
作者 Sudhanshu P Raikwar Apoorva S Raikwar +1 位作者 Shyam S Chaurasia Rajiv R Mohan 《World Journal of Translational Medicine》 2016年第1期1-13,共13页
Gene editing has recently emerged as a promising technology to engineer genetic modifications precisely in the genome to achieve long-term relief from corneal disorders.Recent advances in the molecular biology leading... Gene editing has recently emerged as a promising technology to engineer genetic modifications precisely in the genome to achieve long-term relief from corneal disorders.Recent advances in the molecular biology leading to the development of clustered regularly interspaced short palindromic repeats(CRISPRs) and CRISPR-associated systems,zinc finger nucleases and transcription activator like effector nucleases have ushered in a new era for high throughput in vitro and in vivo genome engineering.Genome editing can be successfully used to decipher complex molecular mechanisms underlying disease pathophysiology,develop innovative next generation gene therapy,stem cell-based regenerative therapy,and personalized medicine for corneal and other ocular diseases.In this review we describe latest developments in the field of genome editing,current challenges,and future prospects for the development of personalized genebased medicine for corneal diseases.The gene editing approach is expected to revolutionize current diagnostic and treatment practices for curing blindness. 展开更多
关键词 ADENO-ASSOCIATED virus Clustered Regularly-Interspaced SHORT Palindromic Repeats associated protein 9 Cornea Clustered regularly interspaced SHORT palindromic repeat double strand breaks GENE EDITING sgRNA GENE targeting Homology directed repair Homologous recombination Indels LENTIVIRAL vector Protospacer-adjacent motif Transcription activator like effector nucleaseS Zinc finger nucleaseS
下载PDF
Genome engineering technologies for targeted genetic modification in plants
9
作者 Wei Tang Anna Y. Tang 《Journal of Forestry Research》 SCIE CAS CSCD 2018年第4期875-887,共13页
Well-established targeted technologies to engi- neer genomes such as zinc-finger nuclease-based editing (ZFN), transcription activator-like effector nuclease-based editing (TALEN), and clustered regularly interspa... Well-established targeted technologies to engi- neer genomes such as zinc-finger nuclease-based editing (ZFN), transcription activator-like effector nuclease-based editing (TALEN), and clustered regularly interspaced short palindromic repeats and associated protein system-based editing (CRISPR/Cas) are proving to advance basic and applied research in numerous plant species. Compared with systems using ZFNs and TALENs, the most recently developed CRISPR/Cas system is more efficient due to its use of an RNA-guided nuclease to generate double-strand DNA breaks. To accelerate the applications of these technologies, we provide here a detailed overview of these systems, highlight the strengths and weaknesses of each, summarize research advances made with these technologies in model and crop plants, and discuss their applications in plant functional genomics. Such targeted approaches for genetically modifying plants will benefit agricultural production in the future. 展开更多
关键词 double-stranded DNA break Genomeediting CRISPR system Transcription activator-likeeffector nucleases Zinc-finger nucleases
下载PDF
双链特异性核酸酶介导的高灵敏度microRNA分析 被引量:3
10
作者 李晓利 王愈聪 +3 位作者 张学晶 赵云颉 刘成辉 李正平 《化学学报》 SCIE CAS CSCD 北大核心 2014年第3期395-400,共6页
基于氧化石墨烯(GO)对荧光标记单链DNA探针的荧光猝灭效应以及双链特异性核酸酶(DSN)选择性切割DNA/RNA杂合结构中DNA单链的特性,本文建立了一种新型恒温信号放大方法用于microRNA(miRNA)的高灵敏度检测.靶标miRNA首先与荧光DNA探针杂交... 基于氧化石墨烯(GO)对荧光标记单链DNA探针的荧光猝灭效应以及双链特异性核酸酶(DSN)选择性切割DNA/RNA杂合结构中DNA单链的特性,本文建立了一种新型恒温信号放大方法用于microRNA(miRNA)的高灵敏度检测.靶标miRNA首先与荧光DNA探针杂交,DSN能够特异性地将杂合双链中的DNA探针水解为碎片但不会降解miRNA,GO对酶切产生的寡核苷酸碎片吸附能力显著降低,使得荧光基团远离GO表面而不被猝灭.释放出的miRNA可再次发生与荧光DNA探针杂交、DSN酶切等反应,如此反复,可实现恒温条件下一个miRNA分子与多个探针杂交、酶切、释放荧光基团的循环过程,最终体系的荧光信号得到显著放大,通过记录体系的荧光信号即可实现对靶标miRNA的灵敏检测. 展开更多
关键词 MICRORNA 双链特异性核酸酶 氧化石墨烯 恒温信号扩增 荧光
原文传递
上一页 1 下一页 到第
使用帮助 返回顶部