Manipulating the Caenorhabditis elegans genome using mariner transposons.
Tc1, one of the founding members of the Tc1/mariner transposon superfamily, was identified in the nematode Caenorhabditis elegans more than 25 years ago. Over the years, Tc1 and other endogenous mariner transposons became valuable tools for mutagenesis and targeted gene inactivation in C. elegans. However, transposition is naturally repressed in the C. elegans germline by an RNAi-like mechanism, necessitating the use of mutant strains in which transposition was globally derepressed, which causes drawbacks such as uncontrolled proliferation of the transposons in the genome and accumulation of background mutations. The more recent mobilization of the Drosophila mariner transposon Mos1 in the C. elegans germline circumvented the problems inherent to endogenous transposons. Mos1 transposition strictly depends on the expression of the Mos transposase, which can be controlled in the germline using inducible promoters. First, Mos1 can be used for insertional mutagenesis. The mobilization of Mos1 copies present on an extrachromosomal array results in the generation of a small number of Mos1 genomic insertions that can be rapidly cloned by inverse PCR. Second, Mos1 insertions can be used for genome engineering. Triggering the excision of a genomic Mos1 insertion causes a chromosomal break, which can be repaired by transgene-instructed gene conversion. This process is used to introduce specific changes in a given gene, such as point mutations, deletions or insertions of a tag, and to create single-copy transgenes.
Robert
Valérie J
VJ
Ecole Normale Supérieure, Biologie Cellulaire de Synapse, Inserm, U789, 75005 Paris, France.
Bessereau
Jean-Louis
JL
D000818:Animals; D017173:Caenorhabditis elegans / Q000235:genetics*; D029742:Caenorhabditis elegans Proteins / Q000235:genetics*; D049109:Cell Proliferation; D004251:DNA Transposable Elements; D004268:DNA-Binding Proteins / Q000235:genetics*; D004330:Drosophila; D017238:Genes, Helminth; D005818:Genetic Engineering; D016678:Genome; D049751:Genome, Helminth; D008954:Models, Biological; D016296:Mutagenesis; D016254:Mutagenesis, Insertional; D018626:Retroelements; D019895:Transposases / Q000235:genetics*
D029742:Caenorhabditis elegans Proteins; D004251:DNA Transposable Elements; D004268:DNA-Binding Proteins; D018626:Retroelements; C111652:mariner transposases; D019895:Transposases