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Full-Text Articles in Life Sciences
A Drosophila Resource Of Transgenic Rnai Lines For Neurogenetics, Quan Jian Ni, Lu Ping Liu, Richard Binari, Robert Hardy, Hye Seok Shim, Amanda Cavallaro, Matthew Booker, Barret D. Pfeiffer, Michele Markstein, Hui Wang, Christians Villalta, Todd R. Laverty, Lizabeth A. Perkins, Norbert Perrimon
A Drosophila Resource Of Transgenic Rnai Lines For Neurogenetics, Quan Jian Ni, Lu Ping Liu, Richard Binari, Robert Hardy, Hye Seok Shim, Amanda Cavallaro, Matthew Booker, Barret D. Pfeiffer, Michele Markstein, Hui Wang, Christians Villalta, Todd R. Laverty, Lizabeth A. Perkins, Norbert Perrimon
Michele Markstein
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 …
Exploiting Position Effects And The Gypsy Retrovirus Insulator To Engineer Precisely Expressed Transgenes, Michele Markstein, Chrysoula Pitsouli, Christians Villalta, Susan E. Celniker, Norbert Perrimon
Exploiting Position Effects And The Gypsy Retrovirus Insulator To Engineer Precisely Expressed Transgenes, Michele Markstein, Chrysoula Pitsouli, Christians Villalta, Susan E. Celniker, Norbert Perrimon
Michele Markstein
A major obstacle to creating precisely expressed transgenes lies in the epigenetic effects of the host chromatin that surrounds them. Here we present a strategy to overcome this problem, employing a Gal4-inducible luciferase assay to systematically quantify position effects of host chromatin and the ability of insulators to counteract these effects at phiC31 integration loci randomly distributed throughout the Drosophila genome. We identify loci that can be exploited to deliver precise doses of transgene expression to specific tissues. Moreover, we uncover a previously unrecognized property of the gypsy retrovirus insulator to boost gene expression to levels severalfold greater than at …
Vector And Parameters For Targeted Transgenic Rna Interference In Drosophila Melanogaster, Jian Quan Ni, Michele Markstein, Richard Binari, Barret Pfeiffer, Lu Ping Liu, Christians Villalta, Matthew Booker, Lizabeth Perkins, Norbert Perrimon
Vector And Parameters For Targeted Transgenic Rna Interference In Drosophila Melanogaster, Jian Quan Ni, Michele Markstein, Richard Binari, Barret Pfeiffer, Lu Ping Liu, Christians Villalta, Matthew Booker, Lizabeth Perkins, Norbert Perrimon
Michele Markstein
The conditional expression of hairpin constructs in Drosophila melanogaster has emerged in recent years as a method of choice in functional genomic studies. To date, upstream activating site–driven RNA interference constructs have been inserted into the genome randomly using P-element–mediated transformation, which can result in false negatives due to variable expression. To avoid this problem, we have developed a transgenic RNA interference vector based on the phiC31 site-specific integration method.