Gene-specific knockouts are a defining technology of reverse genetics, allowing phenotypes to be assigned to any of the thousands of genes identified in genome sequencing projects. For exploring the specific function and expression mechanism of genes, the ability to delete genes to the genome of genetic model organisms is essential.
Fig.1 Schematic of MosDel. (Frøkjaer-Jensen et al., 2010) Mos1-mediated deletions (MosDEL) are based on the Mos1 transposon (Transposons are mobile DNA elements that can jump between chromosomes) to make targeted deletions in the genome. The double-strand break is repaired using injected DNA as a template. The repair can delete up to 25 kb of DNA and simultaneously insert a positive selection marker.
To produce a MosDEL, an injection strain is generated by selecting the gene of interest near the Mos1 transposon and crossing it into the unc-119 mutant background. The injection mix contains three plasmids: (1) Mos1 transposase for excision of the transposon, (2) the unc-119 wild type gene flanked by DNA sequence homologous to the region where the strand break occurs but lacking the gene of interest, for selection and for strand repair, (3) a red fluorescent reporter to mark the extrachromosomal array.
We have developed a well-established platform to generate targeted gene knockouts in C. elegans using MosDel, and the workflow is shown below.
Workflows of C. elegans MosDel service- CD BioSciences
CD BioSciences offers a broad and integrated portfolio of laboratory and manufacturing services, especially for customized C. elegans model service. We have developed a systematic platform of targeting knock-outs of genes in C. elegans using MosDel method which can be optimized to your specifications. Please feel free to contact us if you want to learn more about the service.
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