A versatile strategy for gene trapping and trap conversion in emerging model organisms

被引:23
作者
Kontarakis, Zacharias [1 ]
Pavlopoulos, Anastasios [2 ]
Kiupakis, Alexandros [1 ]
Konstantinides, Nikolaos [1 ]
Douris, Vassilis [1 ]
Averof, Michalis [1 ]
机构
[1] Fdn Res & Technol Hellas, Inst Mol Biol & Biotechnol, GR-70013 Iraklion, Crete, Greece
[2] Univ Cambridge, Dept Zool, Lab Dev & Evolut, Cambridge CB2 3EJ, England
来源
DEVELOPMENT | 2011年 / 138卷 / 12期
关键词
Parhyale; Gene trapping; phi C31 integrase; Regeneration; Transgenesis; iTRAC; CRUSTACEAN PARHYALE-HAWAIENSIS; AMPHIPOD CRUSTACEAN; ENHANCER DETECTION; CELL FATES; INTEGRASE; DROSOPHILA; TRANSFORMATION; TRANSGENESIS; EXPRESSION; ELEMENT;
D O I
10.1242/dev.066324
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
Genetic model organisms such as Drosophila, C. elegans and the mouse provide formidable tools for studying mechanisms of development, physiology and behaviour. Established models alone, however, allow us to survey only a tiny fraction of the morphological and functional diversity present in the animal kingdom. Here, we present iTRAC, a versatile gene-trapping approach that combines the implementation of unbiased genetic screens with the generation of sophisticated genetic tools both in established and emerging model organisms. The approach utilises an exon-trapping transposon vector that carries an integrase docking site, allowing the targeted integration of new constructs into trapped loci. We provide proof of principle for iTRAC in the emerging model crustacean Parhyale hawaiensis: we generate traps that allow specific developmental and physiological processes to be visualised in unparalleled detail, we show that trapped genes can be easily cloned from an unsequenced genome, and we demonstrate targeting of new constructs into a trapped locus. Using this approach, gene traps can serve as platforms for generating diverse reporters, drivers for tissue-specific expression, gene knockdown and other genetic tools not yet imagined.
引用
收藏
页码:2625 / 2630
页数:6
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