学术探索
学术期刊
新闻热点
数据分析
智能评审

In vitro analysis of RNA interference in Drosophila melanogaster

被引：83
作者：
Haley, B [1 ]
Tang, GL [1 ]
Zamore, PD [1 ]
机构：
[1] Univ Massachusetts, Sch Med, Dept Biochem & Mol Pharmacol, Worcester, MA 01605 USA
来源：
METHODS | 2003年 / 30卷 / 04期
关键词：
RNA interference; RNAi; double-stranded RNA; small interfering RNA; siRNA; RNA degradation; posttranscriptional gene silencing; PTGS;
D O I：
10.1016/S1046-2023(03)00052-5
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
Double-stranded RNA (dsRNA) triggers the destruction of mRNA sharing sequence with the dsRNA, a phenomenon termed RNA interference (RNAi). The dsRNA is converted by endonucleolytic cleavage into 21- to 23-nt small interfering RNAs (siRNAs), which direct a multiprotein complex, the RNA-induced silencing complex to cleave RNA complementary to the siRNA. RNAi can be recapitulated in vitro in lysates of syncytial blastoderm Drosophila embryos. These lysates reproduce all of the known steps in the RNAi pathway in flies and mammals. Here we explain how to prepare and use Drosophila embryo lysates to dissect the mechanism of RNAi. (C) 2003 Elsevier Science (USA). All rights reserved.
引用
收藏
页码：330 / 336
页数：7
相关论文
共 27 条
  • [1] Double-stranded RNA specifically disrupts gene expression during planarian regeneration
    Alvarado, AS
    Newmark, PA
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1999, 96 (09) : 5049 - 5054
  • [2] Role for a bidentate ribonuclease in the initiation step of RNA interference
    Bernstein, E
    Caudy, AA
    Hammond, SM
    Hannon, GJ
    [J]. NATURE, 2001, 409 (6818) : 363 - 366
  • [3] Specific interference with gene expression induced by long, double-stranded RNA in mouse embryonal teratocarcinoma cell lines
    Billy, E
    Brondani, V
    Zhang, HD
    Müller, U
    Filipowicz, W
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2001, 98 (25) : 14428 - 14433
  • [4] dsRNA-mediated gene silencing in cultured Drosophila cells:: a tissue culture model for the analysis of RNA interference
    Caplen, NJ
    Fleenor, J
    Fire, A
    Morgan, RA
    [J]. GENE, 2000, 252 (1-2) : 95 - 105
  • [5] Specific inhibition of gene expression by small double-stranded RNAs in invertebrate and vertebrate systems
    Caplen, NJ
    Parrish, S
    Imani, F
    Fire, A
    Morgan, RA
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2001, 98 (17) : 9742 - 9747
  • [6] Use of double-stranded RNA interference in Drosophila cell lines to dissect signal transduction pathways
    Clemens, JC
    Worby, CA
    Simonson-Leff, N
    Muda, M
    Maehama, T
    Hemmings, BA
    Dixon, JE
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2000, 97 (12) : 6499 - 6503
  • [7] RNA interference is mediated by 21-and 22-nucleotide RNAs
    Elbashir, SM
    Lendeckel, W
    Tuschl, T
    [J]. GENES & DEVELOPMENT, 2001, 15 (02) : 188 - 200
  • [8] Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells
    Elbashir, SM
    Harborth, J
    Lendeckel, W
    Yalcin, A
    Weber, K
    Tuschl, T
    [J]. NATURE, 2001, 411 (6836) : 494 - 498
  • [9] Analysis of gene function in somatic mammalian cells using small interfering RNAs
    Elbashir, SM
    Harborth, J
    Weber, K
    Tuschl, T
    [J]. METHODS, 2002, 26 (02) : 199 - 213
  • [10] Functional anatomy of siRNAs for mediating efficient RNAi in Drosophila melanogaster embryo lysate
    Elbashir, SM
    Martinez, J
    Patkaniowska, A
    Lendeckel, W
    Tuschl, T
    [J]. EMBO JOURNAL, 2001, 20 (23) : 6877 - 6888
123