title: Case studies of ends-out gene targeting in Drosophila.
abstract: Ends-in and ends-out gene replacement approaches have been successfully used to disrupt Drosophila genes involved in a variety of biological processes. These methods combine double-strand breaks and homologous recombination to replace a targeted chromosome region with a designed DNA sequence. Unfortunately, these methods require large numbers of single animal crosses, making them both time consuming and labor intensive. Here, we designed a single complete targeting vector for use in a mass crossing ends-out gene targeting study. Importantly, our gene targeting method included a balancer chromosome to block endogenous homologous chromosome pairing and to promote pairing between the foreign targeting DNA fragment and the targeted chromosome. This technique provided successful and efficient gene replacement, greatly facilitating the gene knockout procedure.
authors.affiliation: Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, China.
authors.forename: Haiyang
authors.initials: H
authors.lastname: Chen
authors.forename: Zhiguo
authors.initials: Z
authors.lastname: Ma
authors.forename: Zhonghua
authors.initials: Z
authors.lastname: Liu
authors.forename: Yuan
authors.initials: Y
authors.lastname: Tian
authors.forename: Yanhui
authors.initials: Y
authors.lastname: Xiang
authors.forename: Chao
authors.initials: C
authors.lastname: Wang
authors.forename: Matthew P
authors.initials: MP
authors.lastname: Scott
authors.forename: Xun
authors.initials: X
authors.lastname: Huang
mesh_terms: D000818:Animals; D015139:Blotting, Southern; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D004330:Drosophila / Q000235:genetics*; D055786:Gene Knockout Techniques / Q000379:methods; D018390:Gene Targeting / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D015203:Reproducibility of Results
chemical_list: D004247:DNA