A semi-automated high-throughput approach to the generation of transposon insertion mutants in the nematode Caenorhabditis elegans.
The generation of a large collection of defined transposon insertion mutants is of general interest to the Caenorhabditis elegans research community and has been supported by the European Union. We describe here a semi-automated high-throughput method for mutant production and screening, using the heterologous transposon Mos1. The procedure allows routine culture of several thousand independent nematode strains in parallel for multiple generations before stereotyped molecular analyses. Using this method, we have already generated >17 500 individual strains carrying Mos1 insertions. It could be easily adapted to forward and reverse genetic screens and may influence researchers faced with making a choice of model organism.
Duverger
Yohann
Y
Centre d'Immunologie de Marseille-Luminy, Université de la Méditerranée, Case 906, 13288 Marseille cedex 9, France.
Belougne
Jérôme
J
Scaglione
Sarah
S
Brandli
Dominique
D
Beclin
Christophe
C
Ewbank
Jonathan J
JJ
D000818:Animals; D017173:Caenorhabditis elegans / Q000235:genetics*; D004251:DNA Transposable Elements; D004268:DNA-Binding Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000032:analysis; D049408:Luminescent Agents / Q000032:analysis; D046228:Microarray Analysis; D016254:Mutagenesis, Insertional / Q000379:methods*; D019895:Transposases / Q000032:analysis / Q000235:genetics / Q000378:metabolism*
D004251:DNA Transposable Elements; D004268:DNA-Binding Proteins; D049408:Luminescent Agents; C111652:mariner transposases; D049452:Green Fluorescent Proteins; D019895:Transposases