一种基于DNA载体的RNA干扰技术，用于抑制哺乳动物细胞中的基因表达。
A DNA vector-based RNAi technology to suppress gene expression in mammalian cells.
作者信息
Sui Guangchao, Soohoo Christina, Affar El Bachir, Gay Frédérique, Shi Yujiang, Forrester William C, Shi Yang
机构信息
Department of Pathology, Harvard Medical School, 200 Longwood Avenue, Boston, MA 02115,USA.
出版信息
Proc Natl Acad Sci U S A. 2002 Apr 16;99(8):5515-20. doi: 10.1073/pnas.082117599.
Abstract
Double-stranded RNA-mediated interference (RNAi) has recently emerged as a powerful reverse genetic tool to silence gene expression in multiple organisms including plants, Caenorhabditis elegans, and Drosophila. The discovery that synthetic double-stranded, 21-nt small interfering RNA triggers gene-specific silencing in mammalian cells has further expanded the utility of RNAi into mammalian systems. Here we report a technology that allows synthesis of small interfering RNAs from DNA templates in vivo to efficiently inhibit endogenous gene expression. Significantly, we were able to use this approach to demonstrate, in multiple cell lines, robust inhibition of several endogenous genes of diverse functions. These findings highlight the general utility of this DNA vector-based RNAi technology in suppressing gene expression in mammalian cells.
摘要
双链RNA介导的干扰（RNAi）最近已成为一种强大的反向遗传学工具，可在包括植物、秀丽隐杆线虫和果蝇在内的多种生物体中使基因表达沉默。合成的双链21核苷酸小干扰RNA能在哺乳动物细胞中触发基因特异性沉默这一发现，进一步拓展了RNAi在哺乳动物系统中的应用。在此，我们报告一种技术，该技术可在体内从DNA模板合成小干扰RNA，以有效抑制内源性基因表达。重要的是，我们能够利用这种方法在多个细胞系中证明对多种功能各异的内源性基因有强力抑制作用。这些发现凸显了这种基于DNA载体的RNAi技术在抑制哺乳动物细胞基因表达方面的普遍实用性。
相似文献
1
A DNA vector-based RNAi technology to suppress gene expression in mammalian cells.一种基于DNA载体的RNA干扰技术，用于抑制哺乳动物细胞中的基因表达。
Proc Natl Acad Sci U S A. 2002 Apr 16;99(8):5515-20. doi: 10.1073/pnas.082117599.
2
U6 promoter-driven siRNAs with four uridine 3' overhangs efficiently suppress targeted gene expression in mammalian cells.具有四个尿苷3'突出端的U6启动子驱动的小干扰RNA（siRNA）能有效抑制哺乳动物细胞中的靶向基因表达。
Nat Biotechnol. 2002 May;20(5):497-500. doi: 10.1038/nbt0502-497.
3
Analysis of gene function in somatic mammalian cells using small interfering RNAs.利用小干扰RNA分析哺乳动物体细胞中的基因功能。
Methods. 2002 Feb;26(2):199-213. doi: 10.1016/S1046-2023(02)00023-3.
4
Construction of simple and efficient DNA vector-based short hairpin RNA expression systems for specific gene silencing in mammalian cells.构建用于哺乳动物细胞中特定基因沉默的基于简单高效DNA载体的短发夹RNA表达系统。
Methods Mol Biol. 2007;408:223-41. doi: 10.1007/978-1-59745-547-3_13.
5
Long endogenous dsRNAs can induce complete gene silencing in mammalian cells and primary cultures.长链内源性双链RNA可在哺乳动物细胞和原代培养物中诱导完全的基因沉默。
Oligonucleotides. 2003;13(5):381-92. doi: 10.1089/154545703322617069.
6
[RNA interference directed by small hairpin RNA expressed in COS-7 cells].由在COS-7细胞中表达的小发夹RNA介导的RNA干扰
Yi Chuan Xue Bao. 2003 Apr;30(4):295-300.
7
Enhancement of RNA interference effect in P19 EC cells by an RNA-dependent RNA polymerase.一种依赖RNA的RNA聚合酶增强P19胚胎癌细胞中的RNA干扰效应。
Iran Biomed J. 2009 Jan;13(1):19-25.
8
Development of resistance to RNAi in mammalian cells.哺乳动物细胞中RNA干扰抗性的发展。
Ann N Y Acad Sci. 2005 Nov;1058:105-18. doi: 10.1196/annals.1359.019.
9
Baculovirus-mediated gene silencing in insect cells using intracellularly produced long double-stranded RNA.利用细胞内产生的长双链RNA在昆虫细胞中进行杆状病毒介导的基因沉默。
J Biotechnol. 2007 Feb 1;128(2):226-36. doi: 10.1016/j.jbiotec.2006.09.016. Epub 2006 Nov 16.
10
Transposon-based RNAi delivery system for generating knockdown cell lines.用于生成基因敲低细胞系的基于转座子的RNA干扰递送系统。
Biochem Biophys Res Commun. 2004 Apr 9;316(3):643-50. doi: 10.1016/j.bbrc.2004.02.090.
引用本文的文献
1
Gene therapy then and now: A look back at changes in the field over the past 25 years.彼时与此时的基因疗法：回顾过去25年该领域的变化。
Mol Ther. 2025 May 7;33(5):1889-1902. doi: 10.1016/j.ymthe.2025.02.040. Epub 2025 Feb 28.
2
A Recombinant Lentiviral Vegfr2-Silencing Vector Attenuates Roxarsone-Promoted Growth of Rat Vascular Endothelial Cells and Angiogenesis in Matrigel Plug and B16F10 Xenograft Models.一种重组慢病毒Vegfr2沉默载体可减弱洛克沙胂促进大鼠血管内皮细胞生长以及在基质胶栓和B16F10异种移植模型中的血管生成作用。
Vet Sci. 2024 Sep 24;11(10):451. doi: 10.3390/vetsci11100451.
3
Nanoliposomes as nonviral vectors in cancer gene therapy.纳米脂质体作为癌症基因治疗中的非病毒载体。
MedComm (2020). 2024 Jun 25;5(7):e583. doi: 10.1002/mco2.583. eCollection 2024 Jul.
4
Basic Principles of RNA Interference: Nucleic Acid Types and In Vitro Intracellular Delivery Methods.RNA干扰的基本原理：核酸类型及体外细胞内递送方法
Micromachines (Basel). 2023 Jun 27;14(7):1321. doi: 10.3390/mi14071321.
5
Molecular Characterization of U6 Promoters from Orange-Spotted Grouper (Epinephelus coioides) and Its Application in DNA Vector-Based RNAi Technology.橙点石斑鱼（Epinephelus coioides）U6 启动子的分子特征及其在基于 DNA 载体的 RNAi 技术中的应用。
Mar Biotechnol (NY). 2023 Jun;25(3):388-402. doi: 10.1007/s10126-023-10212-9. Epub 2023 May 8.
6
Extracellular vesicle-based delivery of silencing sequences for the treatment of Machado-Joseph disease/spinocerebellar ataxia type 3.基于细胞外囊泡的沉默序列递送来治疗 Machado-Joseph 病/脊髓小脑共济失调 3 型。
Mol Ther. 2023 May 3;31(5):1275-1292. doi: 10.1016/j.ymthe.2023.04.001. Epub 2023 Apr 6.
7
Systemic RNA Interference Defective (SID) genes modulate dopaminergic neurodegeneration in C. elegans.系统性 RNA 干扰缺陷（SID）基因调节秀丽隐杆线虫多巴胺能神经元退行性变。
PLoS Genet. 2022 Aug 19;18(8):e1010115. doi: 10.1371/journal.pgen.1010115. eCollection 2022 Aug.
8
Transcriptome responses of RNAi-mediated knockdown in at different premolt substages.RNAi 介导的 knockdown 在不同蜕皮亚期的转录组反应。
Front Endocrinol (Lausanne). 2022 Jul 28;13:917088. doi: 10.3389/fendo.2022.917088. eCollection 2022.
9
Deficiency of ribosomal proteins reshapes the transcriptional and translational landscape in human cells.核糖体蛋白缺失重塑了人类细胞的转录和翻译景观。
Nucleic Acids Res. 2022 Jul 8;50(12):6601-6617. doi: 10.1093/nar/gkac053.
10
PolyPurine Reverse Hoogsteen Hairpins Work as RNA Species for Gene Silencing.多聚嘌呤反向 Hoogsteen 发夹可作为 RNA 物种用于基因沉默。
Int J Mol Sci. 2021 Sep 16;22(18):10025. doi: 10.3390/ijms221810025.
本文引用的文献
1
Stable suppression of gene expression by RNAi in mammalian cells.RNA干扰在哺乳动物细胞中对基因表达的稳定抑制作用。
Proc Natl Acad Sci U S A. 2002 Feb 5;99(3):1443-8. doi: 10.1073/pnas.032652399. Epub 2002 Jan 29.
2
Cyclin-dependent kinases.细胞周期蛋白依赖性激酶
Chem Rev. 2001 Aug;101(8):2511-26. doi: 10.1021/cr0001030.
3
Specific interference with gene expression induced by long, double-stranded RNA in mouse embryonal teratocarcinoma cell lines.长双链RNA对小鼠胚胎性癌细胞系基因表达的特异性干扰。
Proc Natl Acad Sci U S A. 2001 Dec 4;98(25):14428-33. doi: 10.1073/pnas.261562698. Epub 2001 Nov 27.
4
The rest is silence.其余的便是寂静。
RNA. 2001 Nov;7(11):1509-21.
5
Dicer functions in RNA interference and in synthesis of small RNA involved in developmental timing in C. elegans.Dicer在RNA干扰以及秀丽隐杆线虫发育时序中参与的小RNA合成过程中发挥作用。
Genes Dev. 2001 Oct 15;15(20):2654-9. doi: 10.1101/gad.927801.
6
RNA interference: listening to the sound of silence.RNA干扰：聆听沉默之声。
Nat Struct Biol. 2001 Sep;8(9):746-50. doi: 10.1038/nsb0901-746.
7
A role for the RNase III enzyme DCR-1 in RNA interference and germ line development in Caenorhabditis elegans.核糖核酸酶III（RNase III）酶DCR-1在秀丽隐杆线虫RNA干扰及生殖系发育中的作用。
Science. 2001 Sep 21;293(5538):2269-71. doi: 10.1126/science.1062039. Epub 2001 Aug 2.
8
Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells.21个核苷酸的RNA双链体在培养的哺乳动物细胞中介导RNA干扰。
Nature. 2001 May 24;411(6836):494-8. doi: 10.1038/35078107.
9
RNA interference--2001.RNA干扰——2001年
Genes Dev. 2001 Mar 1;15(5):485-90. doi: 10.1101/gad.880001.
10
Role for a bidentate ribonuclease in the initiation step of RNA interference.双齿核糖核酸酶在RNA干扰起始步骤中的作用。
Nature. 2001 Jan 18;409(6818):363-6. doi: 10.1038/35053110.
Zotero 插件