Talk:GENETIC TOOLS, TRANSGENESIS, RNAi in different species
We are trying several strategies to generate transgenic animals in other Caenorhabditis species. We want to express transgenes in the germline !
1) We have obtained many transgenic strains of C. briggsae expressing markers in the germline using bombardement of the Cbr-unc-119 mutant (Waterston lab) and the pie-1 promoter. Lines are often very nice, although silencing is stronger than in C. elegans. We need to thaw lines on a regular basis to avoid the total extinction of the signal after several generations.
2) We could perform MosSCI in C. briggsae using a Mos1 insertion line from the Felix lab. So far, only one construct (pie-1::GFP::tubulin) could be integrated and expressed well. All the other genes tested so far (all germline genes) could not recombine ! (we don"t understand why it worked only once, the repair plasmid being the same, the integration site being the same)
3) We tried hard to use antibiotic selection to obtain transgenic lines of C. species 5, C. remanei or C. species 11 but we failed. We have tried to use the C. elegans sur-5 or myo-2 promoters as markers of transgenesis. It could be that the promoter has to be adapted for each species.
4) We recently obtained Mos1 insertions lines in C. remanei MY204. Instead of spending time to built repair plasmids, we are now trying the MiniMos technology both in C. remanei and C. species 11.
5) We have tried to generate a unc-119 mutant in C. species 11 using TALENs: we failed. We have not tried yet to use CRISPR
Delattre lab, Lyon, France http://www.ens-lyon.fr/LBMC/NematodeCell/fr/