Method of Inducible Knockdown of Essential Genes in OSC Cell Culture of Drosophila melanogaster

IF 1.5 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY
S. V. Marfina, E. A. Mikhaleva, N. V. Akulenko, S. S. Ryazansky
{"title":"Method of Inducible Knockdown of Essential Genes in OSC Cell Culture of Drosophila melanogaster","authors":"S. V. Marfina, E. A. Mikhaleva, N. V. Akulenko, S. S. Ryazansky","doi":"10.1134/s0026893324020110","DOIUrl":null,"url":null,"abstract":"<h3 data-test=\"abstract-sub-heading\">Abstract</h3><p>An RNA interference-based method was proposed to achieve an inducible knockdown of genes essential for cell viability. In the method, a genetic cassette in which a copper ion-dependent inducible metallothionein promoter controls expression of a siRNA precursor is inserted into a genomic pre-integrated transgene by CRIPSR/Cas9 technology. The endogenous siRNA source allows the gene knockdown in cell cultures that are refractory to conventional transfection with exogenous siRNA. The efficiency of the method was demonstrated in <i>Drosophila</i> ovarian somatic cell culture (OSC) for two genes that are essential for oogenesis: <i>Cul3</i>, encoding a component of the multiprotein ubiquitin-ligase complex with versatile functions in proteostasis, and <i>cut</i>, encoding a transcription factor regulating differentiation of ovarian follicular cells.</p>","PeriodicalId":18734,"journal":{"name":"Molecular Biology","volume":"174 1","pages":""},"PeriodicalIF":1.5000,"publicationDate":"2024-04-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Molecular Biology","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1134/s0026893324020110","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
引用次数: 0

Abstract

An RNA interference-based method was proposed to achieve an inducible knockdown of genes essential for cell viability. In the method, a genetic cassette in which a copper ion-dependent inducible metallothionein promoter controls expression of a siRNA precursor is inserted into a genomic pre-integrated transgene by CRIPSR/Cas9 technology. The endogenous siRNA source allows the gene knockdown in cell cultures that are refractory to conventional transfection with exogenous siRNA. The efficiency of the method was demonstrated in Drosophila ovarian somatic cell culture (OSC) for two genes that are essential for oogenesis: Cul3, encoding a component of the multiprotein ubiquitin-ligase complex with versatile functions in proteostasis, and cut, encoding a transcription factor regulating differentiation of ovarian follicular cells.

Abstract Image

在黑腹果蝇 OSC 细胞培养中诱导性敲除重要基因的方法
摘要 提出了一种基于 RNA 干扰的方法,以实现对细胞活力至关重要的基因的诱导性敲除。在该方法中,通过 CRIPSR/Cas9 技术将铜离子依赖性诱导型金属硫蛋白启动子控制 siRNA 前体表达的基因盒插入基因组预整合转基因中。这种内源 siRNA 可用于基因敲除细胞培养物,而传统的外源 siRNA 转染方法对这种细胞培养物无效。该方法在果蝇卵巢体细胞培养(OSC)中对两个卵子发生所必需的基因进行了高效验证:Cul3编码多蛋白泛素连接酶复合物的一个成分,在蛋白稳态中具有多种功能;cut编码一个转录因子,调节卵巢滤泡细胞的分化。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
Molecular Biology
Molecular Biology 生物-生化与分子生物学
CiteScore
1.30
自引率
8.30%
发文量
78
审稿时长
3 months
期刊介绍: Molecular Biology is an international peer reviewed journal that covers a wide scope of problems in molecular, cell and computational biology including genomics, proteomics, bioinformatics, molecular virology and immunology, molecular development biology, molecular evolution and related areals. Molecular Biology publishes reviews, experimental and theoretical works. Every year, the journal publishes special issues devoted to most rapidly developing branches of physical-chemical biology and to the most outstanding scientists.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信