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Originally published as Genetics Published Articles Ahead of Print on January 16, 2005.
Genetics, Vol. 169, 1779-1785, March 2005, Copyright © 2005
doi:10.1534/genetics.104.038265
Characterization of Mos1-Mediated Mutagenesis in Caenorhabditis elegans
A Method for the Rapid Identification of Mutated Genes
Daniel C. Williams*,
Thomas Boulin
,1,
Anne-Françoise Ruaud,
Erik M. Jorgensen* and
Jean-Louis Bessereau,2
* Department of Biology, University of Utah, Salt Lake City, Utah 84112
Biologie Cellulaire de la Synapse, INSERM, Ecole Normale Supérieure, 75 005 Paris, France
2 Corresponding author: Biologie Cellulaire de la Synapse, INSERM U 497, Ecole Normale Supérieure, 46 rue d'Ulm, 75 005 Paris, France.
E-mail: jlbesse{at}wotan.ens.fr
Insertional mutagenesis with a heterologous transposon provides a method to rapidly determine the molecular identity of mutated genes. The Drosophila transposon Mos1 can be mobilized to cause mutations in Caenorhabditis elegans (BESSEREAU et al. 2001); however, the mutagenic rate was initially too low for use in most forward genetic screens. To increase the effectiveness of Mos1-mediated mutagenesis we examined the conditions influencing Mos1 transposition. First, optimal transposition occurs 24 hr after expression of the transposase and is unlikely to occur in differentiated sperm or oocytes. Second, transposition is limited to germ-cell nuclei that contain donor elements, but the transposase enzyme can diffuse throughout the gonad syncytium. Third, silencing of transposition is caused by changes in the donor array that occur over time. Finally, multiple transposition events occur in individual germ cells. By using screening techniques based on these results, Mos1 mutagenicity was increased to within an order of magnitude of chemical mutagens.
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