The diplogastrid nematode Pristionchus pacificus is a nematode model system for comparative studies to Caenorhabditis elegans and integrative evolutionary biology aiming for interdisciplinary approaches of evo-devo, population genetics, and ecology. For this, fieldwork can be combined with laboratory studies, and P. pacificus has a welldeveloped methodological toolkit of forward genetics, whole genome sequencing, DNA-mediated transformation, and various –omics platforms. Here, we establish CRISPR/Cas9- based gene inactivation and describe various boundary conditions of this methodology for P. pacificus. Specifically, we demonstrate that most mutations arise within the first 9 hours after injections. We systematically tested the efficiency of sgRNAs targeting different exons in Ppa-dpy-1 and characterized the molecular nature of the induced mutations. Finally, we provide a protocol that might also be useful for researchers working with other non-Caenorhabditis nematodes.
CITATION STYLE
Witte, H., Moreno, E., Rödelsperger, C., Kim, J., Kim, J. S., Streit, A., & Sommer, R. J. (2014). Gene inactivation using the CRISPR/Cas9 systemin the nematode Pristionchus pacificus. Development Genes and Evolution, 225(1), 55–62. https://doi.org/10.1007/s00427-014-0486-8
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