Single-Copy Knock-In Loci for Defined Gene Expression in <i>Caenorhabditis elegans</i>

Abstract

We have generated a single-copy knock-in loci for defined gene expression (SKI LODGE) system to insert any DNA by CRISPR/Cas9 at defined safe harbors in the <i>Caenorhabditis elegans</i> genome. Utilizing a single crRNA guide, which also acts as a Co-CRISPR enrichment marker, any DNA sequence can be introduced as a single copy, regulated by different tissue-specific promoters. The SKI LODGE system provides a fast, economical, and effective approach for generating single-copy ectopic transgenes in <i>C. elegans</i>.