Directed, efficient, and versatile modifications of the Drosophila genome by genomic engineering

• DOI: 10.1073/pnas.0900641106
• Publication Date: 2009-05-09T01:00:09Z
• Authors: Juan Huang; Wenke Zhou; Wei Dong; Annie M. Watson; Yang Hong
• Keywords: Genome Engineering; genomic DNA

With the completion of genome sequences major model organisms, increasingly sophisticated genetic tools are necessary for investigating complex and coordinated functions genes. Here we describe a manipulation system termed "genomic engineering" in Drosophila. Genomic engineering is 2-step process that combines ends-out (replacement) gene targeting with phage integrase phiC31-mediated DNA integration. First, through an improved modified method, founder knock-out line generated by deleting target replacing it integration site phiC31. Second, phiC31 used to reintroduce target-gene into native locus line. permits directed highly efficient modifications chosen genomic virtually any desired mutant allele. We have successfully applied scheme on 6 different genes at their loci more than 70 unique alleles.