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          Institute: MPI für Entwicklungsbiologie     Collection: Abteilung 4 - Evolutionary Biology (R. Sommer)     Display Documents



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ID: 708668.0, MPI für Entwicklungsbiologie / Abteilung 4 - Evolutionary Biology (R. Sommer)
Gene inactivation using the CRISPR/Cas9 system in the nematode Pristionchus pacificus
Authors:Witte, H.; Moreno, E.; Rodelsperger, C.; Kim, J.; Kim, J. S.; Streit, A.; Sommer, R. J.
Language:English
Date of Publication (YYYY-MM-DD):2015-01
Title of Journal:Development Genes and Evolution
Journal Abbrev.:Dev Genes Evol
Volume:225
Issue / Number:1
Start Page:55
End Page:62
Sequence Number of Article:25548084
Review Status:Internal review
Audience:Experts Only
Abstract / Description: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.
External Publication Status:published
Document Type:Article
Communicated by:MPI für Entwickungsbiologie
Affiliations:MPI für Entwicklungsbiologie/Abteilung 4 - Evolutionsbiologie (Ralf J. Sommer)
Identifiers:ISSN:1432-041X (Electronic) 0949-944X (Linking) %R 10.1... [ID No:1]
URL:http://www.ncbi.nlm.nih.gov/pubmed/25548084 [ID No:2]
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