Iterative editing of multiple genes using CRISPR/Cas9 in C. elegans

Certain sets of genes are derived from gene duplication and share substantial sequence similarity in C. elegans , presenting a significant challenge in determining the specific roles of each gene and their collective impact on cellular processes. Here, we show that a collection of genes can be disrupted in a single animal via multiple rounds of CRISPR/Cas9 mediated genome editing. We found that up to three genes can be simultaneously disrupted in a single editing event with high efficiency. Our approach offers an opportunity to explore the genetic interaction and molecular underpinning of gene clusters with redundant function.