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Lookup NU author(s): Dr Hayden Homer, Dr Alexander McDougall, Dr Mark Levasseur, Professor Alison Murdoch, Professor Mary Herbert
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Although female meiosis I errors account for the majority of human aneuploidy, their molecular basis is largely unknown. By elucidating gene function, gene knockdown using RNA interference (RNAi) could shed light on this enigmatic process. In practice, however, the extreme paucity of immature human oocytes makes the evaluation of gene-targeting tools difficult. Here, we undertake RNAi in human oocytes and describe an approach employing mouse oocytes which could overcome the problem of limited biological material. We designed a short interfering RNA (siRNA) designated si539 to target the human mitotic arrest deficient 2 (hMad2) spindle checkpoint component. In human oocytes microinjected with si539, the hMad2 signal detected by Western blotting was 85-92% less intense than in oocytes injected with control siRNA indicating efficient silencing. Further examination of si539's targeting efficiency was undertaken using a green fluorescent protein (GFP)-tagged hMad2 mRNA construct in mouse oocytes. Consistent with Western blot analysis, si539 reduced hMad2-GFP expression in mouse oocytes by ∼94% and relieved the meiosis I arrest otherwise induced by hMad2-GFP in mouse oocytes. By facilitating the investigation of candidate genes involved in regulating human female meiosis I, this approach can bring us closer to understanding the origins of aneuploidies such as Down's syndrome. © The Author 2005. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved.
Author(s): Homer HA, McDougall A, Levasseur M, Murdoch AP, Herbert M
Publication type: Article
Publication status: Published
Journal: Molecular Human Reproduction
Year: 2005
Volume: 11
Issue: 6
Pages: 397-404
Print publication date: 01/06/2005
ISSN (print): 1360-9947
ISSN (electronic): 1460-2407
Publisher: Oxford University Press
URL: http://dx.doi.org/10.1093/molehr/gah184
DOI: 10.1093/molehr/gah184
PubMed id: 15908454
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