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Case studies of ends‐out gene targeting in Drosophila
Author(s) -
Chen Haiyang,
Ma Zhiguo,
Liu Zhonghua,
Tian Yuan,
Xiang Yanhui,
Wang Chao,
Scott Matthew P.,
Huang Xun
Publication year - 2009
Publication title -
genesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.093
H-Index - 110
eISSN - 1526-968X
pISSN - 1526-954X
DOI - 10.1002/dvg.20501
Subject(s) - homologous recombination , gene , biology , genetics , gene targeting , chromosome , homologous chromosome , gene knockout , dna , computational biology
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. genesis 47:305–308, 2009. © 2009 Wiley‐Liss, Inc.