Methods for Studying the Biological Consequences of Endo-siRNA Deficiency in Drosophila melanogaster

  • Do-Hwan Lim
  • Chun-Taek Oh
  • Sung-Jun HanEmail author
  • Young Sik LeeEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1173)


Endogenous small interfering RNAs (endo-siRNAs) are a newly emerged class of small regulatory RNAs. In Drosophila melanogaster, the production of endo-siRNAs depends on the RNase III enzyme Dicer-2 (Dcr-2). Loss of dcr-2 function reduces the resistance of adult flies to various stresses and shortens their life span. The mutants also exhibit alterations in carbohydrate and lipid metabolism. These findings suggest that the endo-siRNA pathway plays a protective role during exposure to stress and aging in D. melanogaster, possibly by regulating metabolic homeostasis. Here, we describe the methods that were used to discover the phenotypes associated with endo-siRNA deficiency by using dcr-2 null mutants.


Drosophila melanogaster Endogenous siRNAs Dicer-2 Stress resistance Life span Metabolism 



This work was supported by a grant from the National Research Foundation of Korea (NRF) funded by the Korea government (Ministry of Education, Science, and Technology [MEST]; 2011-0016863 to Y.S.L.) and the NRF grant funded by MEST (2012-00011, Gyeonggi-do and KISTI to S.J.H.).


  1. 1.
    Kim VN, Han J, Siomi MC (2009) Biogenesis of small RNAs in animals. Nat Rev Mol Cell Biol 10:126–139PubMedCrossRefGoogle Scholar
  2. 2.
    Okamura K, Lai EC (2008) Endogenous small interfering RNAs in animals. Nat Rev Mol Cell Biol 9:673–678PubMedCentralPubMedCrossRefGoogle Scholar
  3. 3.
    Lee YS, Nakahara K, Pham JW, Kim K, He Z, Sontheimer EJ, Carthew RW (2004) Distinct roles for Drosophila Dicer-1 and Dicer-2 in the siRNA/miRNA silencing pathways. Cell 117:69–81PubMedCrossRefGoogle Scholar
  4. 4.
    Lim DH, Oh CT, Lee L, Hong JS, Noh SH, Hwang S, Kim S, Han SJ, Lee YS (2011) The endogenous siRNA pathway in Drosophila impacts stress resistance and lifespan by regulating metabolic homeostasis. FEBS Lett 585:3079–3085PubMedCrossRefGoogle Scholar
  5. 5.
    Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  1. 1.College of Life Sciences and BiotechnologyKorea UniversitySeoulSouth Korea
  2. 2.Drug Biology GroupInstitut Pasteur KoreaSeongnam-SiSouth Korea

Personalised recommendations