RNAi-Mediated Inactivation of Autophagy Genes in Caenorhabditis elegans.

Cold Spring Harbor Protocols
Nicholas J Palmisano, Alicia Meléndez

Abstract

RNA interference (RNAi) is a process that results in the sequence-specific silencing of endogenous mRNA through the introduction of double-stranded RNA (dsRNA). In the nematode Caenorhabditis elegans, RNA inactivation can be used at any specific developmental stage or during adulthood to inhibit a given target gene. Investigators can take advantage of the fact that, in C. elegans, RNAi is unusual in that it is systemic, meaning that dsRNA can spread throughout the animal and can affect virtually all tissues except neurons. Here, we describe a protocol for the most common method to achieve RNAi in C. elegans, which is to feed them bacteria that express dsRNA complementary to a specific target gene. This method has various advantages, including the availability of libraries that essentially cover the whole genome, the ability to treat animals at any developmental stage, and that it is relatively cost effective. We also discuss how RNAi specific to autophagy genes has proven to be an excellent method to study the role of these genes in autophagy, as well as other cellular and developmental processes, while also highlighting the caveats that must be applied.

References

Nov 6, 1998·Nature·L Timmons, A Fire
Mar 31, 2000·Science·A GrishokC C Mello
Feb 9, 2002·Science·William M WinstonCraig P Hunter
Aug 15, 2002·Current Biology : CB·Femke SimmerRonald H A Plasterk
Oct 19, 2004·Genome Research·Jean-François RualMarc Vidal
Jan 6, 2007·Autophagy·Eszter S HarsLeroy F Liu
Jan 11, 2007·Proceedings of the National Academy of Sciences of the United States of America·Caroline SchmitzHarald Hutter
Aug 8, 2007·Gene·Hiroshi QadotaKozo Kaibuchi
Feb 3, 2009·Methods in Enzymology·Alicia MeléndezMalene Hansen
Jun 1, 2010·Nature Methods·Andrea CalixtoMartin Chalfie
Sep 13, 2011·Current Biology : CB·Louis R LapierreMalene Hansen
Jan 6, 2016·Cold Spring Harbor Protocols·Nicholas J Palmisano, Alicia Meléndez
Jan 6, 2016·Cold Spring Harbor Protocols·Nicholas J Palmisano, Alicia Meléndez

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Citations

Feb 3, 2016·Cold Spring Harbor Protocols·Nicholas J Palmisano, Alicia Meléndez

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