Gene inactivation using the CRISPR/Cas9 system in the nematode Pristionchus pacificus

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Title
Gene inactivation using the CRISPR/Cas9 system in the nematode Pristionchus pacificus
Author(s)
Hanh Witte; Eduatdo Moreno; Christian Rodelsperger; Jungeun Kim; Jin Soo Kim; Andrian Streit; Ralf J. Sommer
Publication Date
2015-01
Journal
DEVELOPMENT GENES AND EVOLUTION, v.255, no.1, pp.55 - 62
Publisher
SPRINGER
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 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.
URI
https://pr.ibs.re.kr/handle/8788114/822
ISSN
0949-944X
Appears in Collections:
Center for Genome Engineering(유전체 교정 연구단) > Journal Papers (저널논문)
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