How to knock-in genes with multi-epitope tagging in C. elegans using CRISPR/Cas9 gene editing technique?

09 October 2024 0 7K Report

I am interested in inserting a gRNA into the genome of C. elegans and tagging multiple epitopes at both the prodomain and mature domain regions of the target signal sequence to observe the phenotypic effects induced by the regulation of the target protein. Currently, I am using CHOPCHOP to design the gRNA and homology arms.

1) What conditions need to be followed when designing the gRNA for a knock-in experiment?

2) Additionally, does anyone have suggestions for free tools that can help align and visualize epitopes, gRNA, and restriction sites in a single frame?

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