From the Cover: Directed, efficient, and versatile modifications of the Drosophila genome by genomic engineering.

Abstract:

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

authors

Huang J,Zhou W,Dong W,Watson AM,Hong Y

doi

10.1073/pnas.0900641106

subject

Has Abstract

pub_date

2009-05-19 00:00:00

pages

8284-9

issue

20

eissn

0027-8424

issn

1091-6490

pii

0900641106

journal_volume

106

pub_type

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