您好，欢迎访问湖北省农业科学院 机构知识库！
湖北省农业科学院机构知识库

全部

  • 全部
  • 标题
  • 学者
  • 机构
  • 关键词

Construction and application of a built-in dual luciferase reporter for microRNA functional analysis

文献类型： 外文期刊

作者： Bi, Yanzhen 1 ; Zheng, Xinmin 1 ; Shao, Changwei 2 ; Pan, Wen 3 ; Jiang, Li 2 ; Ouyang, Huiwu 2 ;

作者机构： 1.Hubei Acad Agr Sci, Inst Anim Husb & Vet, Hubei Key Lab Anim Embryo Engn & Mol Breeding, Wuhan, Peoples R China

2.Wuhan Univ, Coll Life Sci, RNA Grp, Wuhan, Peoples R China

3.Sun Yat Sen Univ, Affiliated Hosp 3, Guangzhou, Guangdong, Peoples R China

关键词： biosensor;luciferase;ligase-independent;miRNA;target

期刊名称：ELECTRONIC JOURNAL OF BIOTECHNOLOGY （ 影响因子：2.8； 五年影响因子：3.379 ）

ISSN： 0717-3458

年卷期： 2011 年 14 卷 2 期

页码：

收录情况： SCI

摘要： Background: As key gene regulators, microRNAs post-transcriptionally modulate gene expression via binding to partially complementary sequence in the 3' UTR of target mRNA. An accurate, rapid and quantitative tool for sensing and validation of miRNA targets is of crucial significance to decipher the functional implications of miRNAs in cellular pathways. Results: Taking advantage of an improved restriction-free cloning method, we engineered a novel built-in dual luciferase reporter plasmid where Firefly and Renilla luciferase genes were assembled in a single plasmid named "pFila". This design eliminates the transfection of a separate control plasmid and thus minimizes the time and labor required for miRNA-target sensing assays. pFila consistently produces Firefly and Renilla luciferase activities when transfected into human-, monkey-and mouse-derived mammalian cell systems. Moreover, pFila is capable of recapitulating the interaction of miR-16 and its known target CCNE1 in Hela cells. Additionally, pFila is shown to be a sensitive miR-biosensor by evaluating the inhibition efficiency of endogenous miRNA. Conclusions: pFila would facilitate miRNA target identification and verification in a rapid and simplified manner. Also, pFila is a sensitive biosensor for active miRNA profiling in vivo.

  • 相关文献

[1]miRNA调控家禽卵泡发育的研究进展. 吴艳,皮劲松,潘爱銮,蒲跃进,申杰,张昊,梁振华,孙静,杜金平. 2018

[2]调控CD163基因表达的miRNA鉴定及其在PRRSV感染中的作用分析. 吴俊静,乔木,武华玉,彭先文,梅书棋. 2017

[3]Ssc-miR-27b-3p在猪繁殖与呼吸综合征病毒感染中的作用分析. 吴俊静,季子云,乔木,武华玉,刘贵生,彭先文,梅书棋. 2018

[4]circ-13267通过let-7-19/ERBB4通路调控蛋鸭卵泡颗粒细胞凋亡. 吴艳,张昊,梁振华,潘爱銮,申杰,蒲跃进,黄涛,皮劲松,杜金平. 2022

[5]利用双荧光素酶报告系统鉴定靶向作用猪CD163基因的miRNA. 吴俊静,彭先文,乔木,武华玉,刘贵生,梅书棋. 2015

[6]基于多工具的山羊circRNA-miRNA靶向关系分析. 陶虎,杨娟,许铭洙,刘泽林,张年,刘洋,陈明新. 2021

[7]猪肌内脂肪前体细胞成脂分化中miRNA差异分析. 张新雨,杨涛,吴俊静,乔木,徐忠,张宇,冯越,李梓芃,彭先文,梅书棋,李凤娥,周佳伟. 2024

作者其他论文 更多>>
置顶
购物车
反馈

© 京ICP备09089781号-29 主办单位：湖北省农业科学院

学术资源： 中国农业科学院 农业专业知识服务系统 农科联盟资源共建共享平台 中国农业科技文献与信息服务平台 中国农业期刊集成服务平台