SKI LODGE
Overview
The SKI LODGE cassettes were introduced into intergenic regions that are known to give stable expression.
The SKI LODGE system allows insertion of single-copy transgenes in C. elegans into safe harbor loci using CRISPR/Cas9 editing. Utilizing the dpy-10 crRNA guide both for knock-in and Co-CRISPR edits, the gene of interest can be introduced as a single copy, regulated by different tissue-specific promoters.
Summarized Protocol
Select the SKI LODGE strain to express the gene of interest.
Design primers with homologous recombination arms (~35 bp) for the SKI LODGE cassette (review sequences in Step by Step guide and Strains sections).
Assembly of CRISPR/Cas9 complex in vitro.
Injection.
3-4 days after injection screen for dumpy/rollers worms.
Single out worms and look for fluorescent expression or genotype them.
Outcrossing.