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Large-scale Gene Knockdown in C. elegans Using dsRNA Feeding Libraries to Generate Robust Loss-of-function Phenotypes
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Large-scale Gene Knockdown in C. elegans Using dsRNA Feeding Libraries to Generate Robust Loss-of-function Phenotypes

Large-scale Gene Knockdown in C. elegans Using dsRNA Feeding Libraries to Generate Robust Loss-of-function Phenotypes

DOI:
10.3791/50693-v

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18:38 min

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September 25, 2013

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, ,
1Molecular and Cellular Biology Program,University of Massachusetts, Amherst, 2Neuroscience and Behavior Program,University of Massachusetts, Amherst, 3Department of Biochemistry and Molecular Biology,University of Massachusetts, Amherst

Chapters

  • 00:05Title
  • 02:54How to Determine Plasmid Loss
  • 04:32Making Temporary Copies of the Library Omniplates
  • 06:35Preparation of Bacteria for Feeding Worms
  • 09:28Determining the Concentration of L1 Arrested C. elegans Larvae
  • 10:53dsRNA Feeding Screen
  • 12:37Results: Effects of egl-30, dpy-17, pat-10, and unc-4 Knockdown
  • 17:50Conclusion

Summary

Automatic Translation

Automatic Translation

While dsRNA feeding in C. elegans is a powerful tool to assess gene function, current protocols for large scale feeding screens result in variable knockdown efficiencies. We describe an improved protocol for performing large scale RNAi feeding screens that results in highly efficacious and reproducible knockdown of gene expression.

Tags

Keywords: RNA InterferenceRNAiDsRNA FeedingC. ElegansGene KnockdownLoss-of-function PhenotypesHigh-throughput ScreeningPlasmid LossProtocolNeurons

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