A Drosophila Resource of Transgenic RNAi Lines for Neurogenetics
View/ Open
74481 GEN18241089.pdf (2.825Mb)
Access Status
Full text of the requested work is not available in DASH at this time ("dark deposit"). For more information on dark deposits, see our FAQ.Author
Ni, Jian-Quan
Liu, Lu-Ping
Binari, Richard
Hardy, Robert
Shim, Hye-Seok
Cavallaro, Amanda
Booker, Matthew
Pfeiffer, Barret D.
Markstein, Michele
Wang, Hui
Villalta, Christians
Laverty, Todd R.
Perkins, Lizabeth A.
Perrimon, Norbert
Published Version
https://doi.org/10.1534/genetics.109.103630Metadata
Show full item recordCitation
Ni, Jian-Quan, Lu-Ping Liu, Richard Binari, Robert Hardy, Hye-Seok Shim, Amanda Cavallaro, Matthew Booker, et al. 2009. “A Drosophila Resource of Transgenic RNAi Lines for Neurogenetics.” Genetics 182 (4): 1089–1100. https://doi.org/10.1534/genetics.109.103630.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 h,51)70 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 generate 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.Citable link to this page
http://nrs.harvard.edu/urn-3:HUL.InstRepos:41384502
Collections
- HMS Scholarly Articles [18124]
Contact administrator regarding this item (to report mistakes or request changes)