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MISSION esiRNA for RNAi Screening in Mammalian Cells
JoVE Journal
Biologia
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JoVE Journal Biologia
MISSION esiRNA for RNAi Screening in Mammalian Cells

MISSION esiRNA for RNAi Screening in Mammalian Cells

DOI:
10.3791/2008-v

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15:31 min

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May 12, 2010

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,
1Max Planck Institute of Molecular Cell Biology and Genetics

Capítulos

  • 00:00Título
  • 01:43Introduction
  • 02:22Use of High-Quality MISSION esiRNA Libraries for RNAi Screening
  • 03:49Optimizing Transfection Prior to Screening
  • 05:28Setting Up the Assay and Preparing for the Primary Screen
  • 09:34The Primary Screen
  • 12:03Secondary Screen of the Selected Genes for Hit Validation
  • 13:40Representative Results Screening for Mammalian Cell Cycle Genes Using MISSION esiRNA
  • 14:43Conclusion

Summary

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Tadução automática

Here we use a human esiRNA library in a high-throughput screen for genes involved in cell division. We demonstrate how to set up and conduct an esiRNA screens, as well as how to analyze and validate the results.

Tags

MISSION EsiRNARNAi ScreeningMammalian CellsGene Expression ControlSmall Interfering RNAs (siRNAs)DicerRNA-induced Silencing Complex (RISC)Target MRNA CleavageLoss Of Gene Function StudiesChemical Synthesis Of SiRNAsEndonucleolytic CleavageTarget-specific Long Double-stranded RNAsRNase IIIEndoribonuclease-prepared SiRNAEfficacy Of SiRNAs And EsiRNAsOff-target Effects

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