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

PloS One
Wenke ZhouYang Hong

Abstract

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.

References

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Feb 18, 2003·Proceedings of the National Academy of Sciences of the United States of America·Wei J Gong, Kent G Golic
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Sep 2, 2008·Genetics·Juan HuangYang Hong
Sep 6, 2008·Proceedings of the National Academy of Sciences of the United States of America·Guanjun GaoYikang S Rong
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Apr 4, 2009·Science·Ching Man ChoiBassem A Hassan
May 12, 2009·Proceedings of the National Academy of Sciences of the United States of America·Juan HuangYang Hong
Oct 14, 2009·Fly·Juan HuangYang Hong

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