Abstract
Methods that use homologous recombination to engineer the genome of C. elegans commonly use strains carrying specific insertions of the heterologous transposon Mos1. A large collection of known Mos1 insertion alleles would therefore be of general interest to the C. elegans research community. We describe here the optimization of a semi-automated methodology for the construction of a substantial collection of Mos1 insertion mutant strains. At peak production, more than 5,000 strains were generated per month. These strains were then subject to molecular analysis, and more than 13,300 Mos1 insertions characterized. In addition to targeting directly more than 4,700 genes, these alleles represent the potential starting point for the engineered deletion of essentially all C. elegans genes and the modification of more than 40% of them. This collection of mutants, generated under the auspices of the European NEMAGENETAG consortium, is publicly available and represents an important research resource.
Highlights
The nematode worm Caenorhabditis elegans has long been a model of choice for many areas of biological research because of its powerful genetics
These breaks can be repaired through homologous recombination, using designed transgenic templates, with homology arms that match the genomic sequence on either side of the Mos1 transposon insertion site
We evaluated the number of new Mos1 insertions that fell into the coding regions of exons, which could provoke a loss of gene function
Summary
The nematode worm Caenorhabditis elegans has long been a model of choice for many areas of biological research because of its powerful genetics. Two techniques have become popular, MosTIC, for Mos excisioninduced transgene-instructed gene conversion, [1], and MosSCI, for Mos1-mediated single-copy insertion [2]. Both methods rely on the availability of C. elegans strains carrying integrated copies of the heterologous Mos transposon at defined genomic addresses. Chromosomal breaks can be generated at a single locus through the controlled excision of the Mos transposon These breaks can be repaired through homologous recombination, using designed transgenic templates, with homology arms that match the genomic sequence on either side of the Mos transposon insertion site. A third technique, MosDEL, can be used to generate Mos1-mediated targeted deletions of up to 25 kb [4], allowing null alleles to be generated with relative ease
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
