Characterization of Mos1-mediated mutagenesis in Caenorhabditis elegans: a method for the rapid identification of mutated genes.
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.
Williams
Daniel C
DC
Department of Biology, University of Utah, Salt Lake City, 84112, USA. jlbesse@wotan.ens.fr
Boulin
Thomas
T
Ruaud
Anne-Françoise
AF
Jorgensen
Erik M
EM
Bessereau
Jean-Louis
JL
D000818:Animals; D017173:Caenorhabditis elegans / Q000235:genetics*; D004251:DNA Transposable Elements; D004268:DNA-Binding Proteins / Q000235:genetics; D020868:Gene Silencing; D008957:Models, Genetic; D016296:Mutagenesis; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis; D019895:Transposases
D004251:DNA Transposable Elements; D004268:DNA-Binding Proteins; C111652:mariner transposases; D019895:Transposases