Construction of Transgenic Drosophila by Using the Site-Specific Integrase From Phage C31

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ID: 298486
2004
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Abstract
The phiC31 integrase functions efficiently in vitro and in Escherichia coli, yeast, and mammalian cells, mediating unidirectional site-specific recombination between its attB and attP recognition sites. Here we show that this site-specific integration system also functions efficiently in Drosophila melanogaster in cultured cells and in embryos. Intramolecular recombination in S2 cells on transfected plasmid DNA carrying the attB and attP recognition sites occurred at a frequency of 47%. In addition, several endogenous pseudo attP sites were identified in the fly genome that were recognized by the integrase and used as substrates for integration in S2 cells. Two lines of Drosophila were created by integrating an attP site into the genome with a P element. phiC31 integrase injected into embryos as mRNA functioned to promote integration of an attB-containing plasmid into the attP site, resulting in up to 55% of fertile adults producing transgenic offspring. A total of 100% of these progeny carried a precise integration event at the genomic attP site. These experiments demonstrate the potential for precise genetic engineering of the Drosophila genome with the phiC31 integrase system and will likely benefit research in Drosophila and other insects.
Reference Key
openalex_W2104582907 Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors Amy C. Groth, Matthew Fish, Roel Nusse, Michèle P. Calos
Journal current genetics
Year 2004
DOI
10.1534/genetics.166.4.1775
URL
https://doi.org/10.1534/genetics.166.4.1775
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