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A Drosophila resource of transgenic RNAi lines for neurogenetics
Genetics (2009)
  • Quan Jian Ni
  • Lu Ping Liu
  • Richard Binari
  • Robert Hardy
  • Hye Seok Shim
  • Amanda Cavallaro
  • Matthew Booker
  • Barret D. Pfeiffer
  • Michele Markstein, University of Massachusetts - Amherst
  • Hui Wang
  • Christians Villalta
  • Todd R. Laverty
  • Lizabeth A. Perkins
  • Norbert Perrimon

Conditional expression of hairpin constructs in Drosophila is a powerful method to disrupt the activity of single genes with a spatial and temporal resolution that is impossible, or exceedingly difficult, using classical genetic methods. We previously described a method (Ni et al. 2008) whereby RNAi constructs are targeted into the genome by the phiC31-mediated integration approach using Vermilion-AttB-Loxp-Intron-UAS-MCS (VALIUM), a vector that contains vermilion as a selectable marker, an attB sequence to allow for phiC31-targeted integration at genomic attP landing sites, two pentamers of UAS, the hsp70 core promoter, a multiple cloning site, and two introns. As the level of gene activity knockdown associated with transgenic RNAi depends on the level of expression of the hairpin constructs, we generated a number of derivatives of our initial vector, called the “VALIUM” series, to improve the efficiency of the method. Here, we report the results from the systematic analysis of these derivatives and characterize VALIUM10 as the most optimal vector of this series. A critical feature of VALIUM10 is the presence of gypsy insulator sequences that boost dramatically the level of knockdown. We document the efficacy of VALIUM as a vector to analyze the phenotype of genes expressed in the nervous system and have generated a library of 2282 constructs targeting 2043 genes that will be particularly useful for studies of the nervous system as they target, in particular, transcription factors, ion channels, and transporters.

  • AttP sites,
  • UAS-Luciferase,
  • Gypsy Insulator,
  • Notch RNAi
Publication Date
June 1, 2009
Publisher Statement
doi: 10.1534/genetics.109.103630
Citation Information
Quan Jian Ni, Lu Ping Liu, Richard Binari, Robert Hardy, et al.. "A Drosophila resource of transgenic RNAi lines for neurogenetics" Genetics Vol. 182 Iss. 4 (2009)
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