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The SKI LODGE cassettes were introduced into intergenic regions that are known to give stable expression.

Figure 2

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

  1. Select the SKI LODGE strain to express the gene of interest.

  2. Design primers with homologous recombination arms (~35 bp) for the SKI LODGE cassette (review sequences in Step by Step guide and Strains sections).

  3. Assembly of CRISPR/Cas9 complex in vitro.

  4. Injection.

  5. 3-4 days after injection screen for dumpy/rollers worms.

  6. Single out worms and look for fluorescent expression or genotype them.

  7. Outcrossing.