Biol Methods Protoc 2020 10;5(1):bpaa003. Epub 2020 Feb 10.
Department of Biology, McMaster University, 1280 Main Street West, Hamilton, ON L8S-4K1, Canada.
The CRISPR/Cas system has recently emerged as a powerful tool to engineer the genome of an organism. The system is adopted from bacteria where it confers immunity against invading foreign DNA. This work reports the first successful use of the CRISPR/Cas system in (a cousin of the well-known nematode ), to generate mutations via non-homologous end joining. We recovered deletion alleles of several conserved genes by microinjecting plasmids that express Cas9 endonuclease and an engineered CRISPR RNA corresponding to the DNA sequence to be cleaved. Evidence for somatic mutations and off-target mutations are also reported. Our approach allows for the generation of loss-of-function mutations in genes thereby facilitating a comparative study of gene function.