Screening for gene function in chicken embryo using RNAi and electroporation

被引:140
作者
Pekarik, V [1 ]
Bourikas, D [1 ]
Miglino, N [1 ]
Joset, P [1 ]
Preiswerk, S [1 ]
Stoeckli, ET [1 ]
机构
[1] Univ Zurich, Inst Zool, CH-8057 Zurich, Switzerland
关键词
D O I
10.1038/nbt770
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
In the postgenomic era the elucidation of the physiological function of genes has become the rate-limiting step in the quest to understand the development and function of living organisms. Gene functions cannot be determined by high-throughput methods but require analysis in the context of the entire organism. This is particularly true in the developing vertebrate nervous system(1). Because of its easy accessibility in the egg, the chicken embryo has been the model of choice for developmental in vivo studies. However, its usefulness has been hampered by a lack of methods for genetic manipulation. Here we describe an approach that could compensate for this disadvantage. By combining gene silencing by dsRNA(2) (through RNA interference, RNAi) with in ovo electroporation(3,4), we developed an efficient method to induce loss of gene function in vivo during the development of the chicken CNS. This method opens new possibilities for studying gene function not only by gain-of-function but also by loss-of-function approaches and therefore represents a new tool for functional genomics.
引用
收藏
页码:93 / 96
页数:4
相关论文
共 20 条
  • [1] F-spondin is required for accurate pathfinding of commissural axons at the floor plate
    Burstyn-Cohen, T
    Tzarfaty, V
    Frumkin, A
    Feinstein, Y
    Stoeckli, E
    Klar, A
    [J]. NEURON, 1999, 23 (02) : 233 - 246
  • [2] Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans
    Fire, A
    Xu, SQ
    Montgomery, MK
    Kostas, SA
    Driver, SE
    Mello, CC
    [J]. NATURE, 1998, 391 (6669) : 806 - 811
  • [3] GEFs, GAPs, GDIs and effectors: taking a closer (3D) look at the regulation of Ras-related GTP binding proteins
    Geyer, M
    Wittinghofer, A
    [J]. CURRENT OPINION IN STRUCTURAL BIOLOGY, 1997, 7 (06) : 786 - 792
  • [4] Ikonomov OC, 1998, J NEUROSCI, V18, P6331
  • [5] 'Shocking' developments in chick embryology:: electroporation and in ovo gene expression
    Itasaki, N
    Bel-Vialar, S
    Krumlauf, R
    [J]. NATURE CELL BIOLOGY, 1999, 1 (08) : E203 - E207
  • [6] Double-stranded RNA injection produces null phenotypes in zebrafish
    Li, YX
    Farrell, MJ
    Liu, RP
    Mohanty, N
    Kirby, ML
    [J]. DEVELOPMENTAL BIOLOGY, 2000, 217 (02) : 394 - 405
  • [7] Challenges for neuroscience in a post-genome world
    Lo, DC
    [J]. NATURE NEUROSCIENCE, 2001, 4 (Suppl 11) : 1153 - 1154
  • [8] Momose T, 1999, DEV GROWTH DIFFER, V41, P335
  • [9] Ten years of gene targeting:: targeted mouse mutants, from vector design to phenotype analysis
    Müller, U
    [J]. MECHANISMS OF DEVELOPMENT, 1999, 82 (1-2) : 3 - 21
  • [10] Comparison of three nonviral transfection methods for foreign gene expression in early chicken embryos in ovo
    Muramatsu, T
    Mizutani, Y
    Ohmori, Y
    Okumura, J
    [J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1997, 230 (02) : 376 - 380