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siRNA的筛选识别泛素和泛素样系统压力表生物学途径在培养的哺乳动物细胞
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JoVE Journal Biology
siRNA Screening to Identify Ubiquitin and Ubiquitin-like System Regulators of Biological Pathways in Cultured Mammalian Cells
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siRNA的筛选识别泛素和泛素样系统压力表生物学途径在培养的哺乳动物细胞

DOI:
10.3791/51572-v

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10:43 min

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May 24, 2014

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, , , ,
1MRC Protein Phosphorylation and Ubiquitylation Unit, College of Life Sciences,University of Dundee, 2Centre for Gene Regulation and Expression, College of Life Sciences,University of Dundee

Chapters

  • 00:05Title
  • 01:22Characterizing Hypoxia-responsiveness to Develop the Assay
  • 04:18Primary Screen: First Day
  • 07:27Primary Screen: Subsequent Days
  • 08:27Sample Screen
  • 10:09Conclusion

Summary

Automatic Translation

Automatic Translation

在这里，我们描述了一种方法来执行对象的siRNA“ubiquitome”屏幕来识别对缺氧的HIF1A介导的细胞应答的新型泛素和泛素样调节剂。这可以适用于任何生物学途径，其中一个强大的读出报告基因活性的是可用的。

Tags

SiRNA ScreeningUbiquitinUbiquitin-like SystemBiological PathwaysMammalian CellsPost-translational ModificationUbiquitomeHIF1AHypoxia ResponseTranscription-based Luciferase ReporterAssay DevelopmentPrimary ScreeningSecondary ScreeningTertiary/deconvolution ScreeningTargeted SiRNA LibraryOff-target Effects

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