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W::Neo: A novel dual-selection marker for high efficiency gene targeting in Drosophila

Zhou, W and Huang, J and Watson, AM and Hong, Y (2012) W::Neo: A novel dual-selection marker for high efficiency gene targeting in Drosophila. PLoS ONE, 7 (2).

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We have recently developed a so-called genomic engineering approach that allows for directed, efficient and versatile modifications of Drosophila genome by combining the homologous recombination (HR)-based gene targeting with site-specific DNA integration. In genomic engineering and several similar approaches, a "founder" knock-out line must be generated first through HR-based gene targeting, which can still be a potentially time and resource intensive process. To significantly improve the efficiency and success rate of HR-based gene targeting in Drosophila, we have generated a new dual-selection marker termed W::Neo, which is a direct fusion between proteins of eye color marker White (W) and neomycin resistance (Neo). In HR-based gene targeting experiments, mutants carrying W::Neo as the selection marker can be enriched as much as fifty times by taking advantage of the antibiotic selection in Drosophila larvae. We have successfully carried out three independent gene targeting experiments using the W::Neo to generate genomic engineering founder knock-out lines in Drosophila. © 2012 Zhou et al.


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Item Type: Article
Status: Published
CreatorsEmailPitt UsernameORCID
Zhou, W
Huang, J
Watson, AM
Hong, Yyhong@pitt.eduYHONG
ContributionContributors NameEmailPitt UsernameORCID
Date: 13 February 2012
Date Type: Publication
Journal or Publication Title: PLoS ONE
Volume: 7
Number: 2
DOI or Unique Handle: 10.1371/journal.pone.0031997
Schools and Programs: School of Medicine > Cell Biology and Molecular Physiology
Refereed: Yes
MeSH Headings: Animals; Animals, Genetically Modified; Drosophila--genetics; Eye Proteins--genetics; Gene Targeting--methods; Genetic Engineering--methods; Genetic Markers; Homologous Recombination
Other ID: NLM PMC3278458
PubMed Central ID: PMC3278458
PubMed ID: 22348139
Date Deposited: 12 Sep 2012 21:42
Last Modified: 05 Feb 2019 15:55


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