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Citation
Ni, J.Q., Liu, L.P., Binari, R., Hardy, R., Shim, H.S., Cavallaro, A., Booker, M., Pfeiffer, B.D., Markstein, M., Wang, H., Villalta, C., Laverty, T.R., Perkins, L.A., Perrimon, N. (2009). A Drosophila Resource of Transgenic RNAi Lines for Neurogenetics.  Genetics 182(4): 1089--1100.
FlyBase ID
FBrf0208670
Publication Type
Research paper
Abstract

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.

PubMed ID
PubMed Central ID
PMC2728850 (PMC) (EuropePMC)
Related Publication(s)
Personal communication to FlyBase

Update to TRiP stock collection.
Perkins et al., 2009.8.10, Update to TRiP stock collection. [FBrf0208863]

Update to TRiP stock collection.
Perkins et al., 2009, Update to TRiP stock collection. [FBrf0208864]

Flanking sequence of su(Hw)attP docking sites.
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Secondary IDs
    Language of Publication
    English
    Additional Languages of Abstract
    Parent Publication
    Publication Type
    Journal
    Abbreviation
    Genetics
    Title
    Genetics
    Publication Year
    1916-
    ISBN/ISSN
    0016-6731
    Data From Reference
    Alleles (59)
    Genes (26)
    Datasets (2)
    Molecular Constructs (3)
    Natural transposons (2)
    Insertions (5)
    Experimental Tools (4)
    Transgenic Constructs (52)