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

Kathryn N. Maher; Mary Catanese; Daniel L. Chase

doi:10.3791/50693

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Storstilet Gene Knockdown i C. elegans Brug dsRNA Fodring Biblioteker at skabe en robust tab af funktion fænotyper

JoVE Journal
Biologia

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JoVE Journal Biologia

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

Storstilet Gene Knockdown i C. elegans Brug dsRNA Fodring Biblioteker at skabe en robust tab af funktion fænotyper

DOI:

10.3791/50693-v

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18:38 min

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September 25, 2013

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Kathryn N. Maher, Mary Catanese, Daniel L. Chase

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

Capítulos

00:05Título
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

Tadução automática

Mens dsRNA fodring i C. elegans er et kraftfuldt værktøj til at vurdere gen funktion, nuværende protokoller for storstilede fodring skærme resulterer i variable knockdown effektivitetsgevinster. Vi beskriver en forbedret protokol til at udføre storstilet RNAi fodring skærme, der resulterer i meget effektive og reproducerbar knockdown af genekspression.

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