express restriction enzyme in worms?

Model Organism: Worms Scientific Discussion
1

Hi,

I am wondering whether some people have tried to express restriction enzymes and their target sites in worms? Can I get the transgenic lines somewhere?

Thanks so much.

shouhong

2

Dana Caroll’s lab at Utah have been expressing so-called Zinc Finger Nucleases in worms. These are a fusion of the nuclease domain of the Type IIs enzyme FokI with a three-fingered DNA recognition domain of choice. In pairs these can be made to target very specific sites. They succeeded in getting mobility in the soma for a single site in the genome as evidenced by mis-repair of the anticipated lesions, but I don’t know about the germline. I’d think expression of other enzymes (e.g. EcoRI), would be lethal.

Check Morton et al. 2006, PNAS 103(44):16370-5, “Induction and repair of zinc-finger nuclease-targeted double-strand breaks in Caenorhabditis elegans somatic cells” and also Carroll et al. 2008, Methods Mol. Biol. 435:63-77, “Gene targeting in Drosophila and Caenorhabditis elegans with zinc-finger nucleases.”

Other labs have been expressing recombinases and transposases which also digest DNA at specific sites, but I’m not sure that’s what you had in mind.

3

Thanks very much.