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- Title
Gene inactivation using the CRISPR/Cas9 system in the nematode Pristionchus pacificus.
- Authors
Witte, Hanh; Moreno, Eduardo; Rödelsperger, Christian; Kim, Jungeun; Kim, Jin-Soo; Streit, Adrian; Sommer, Ralf
- Abstract
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 well-developed 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.
- Subjects
GENE silencing; NEMATODE genetics; DIPLOGASTERIDA; COMPARATIVE studies; BIOLOGICAL evolution; POPULATION genetics; GENE targeting
- Publication
Development Genes & Evolution, 2015, Vol 225, Issue 1, p55
- ISSN
0949-944X
- Publication type
Article
- DOI
10.1007/s00427-014-0486-8