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

Witte, H., Moreno, E., Rödelsperger, C., Kim, J., Kim, J.-S., Streit, A., et al. (2015). Gene inactivation using the CRISPR/Cas9 system in the nematode Pristionchus pacificus. Development Genes and Evolution, 225(1), 55-62. doi:10.1007/s00427-014-0486-8.

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Witte, H1, Autor           
Moreno, E1, Autor           
Rödelsperger, C1, Autor           
Kim, J, Autor
Kim, J-S, Autor
Streit, A1, Autor           
Sommer, RJ1, Autor           
Affiliations:
1Department Integrative Evolutionary Biology, Max Planck Institute for Developmental Biology, Max Planck Society, ou_3375786              

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 Zusammenfassung: 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.

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 Datum: 2015-01
 Publikationsstatus: Erschienen
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 Identifikatoren: DOI: 10.1007/s00427-014-0486-8
PMID: 25548084
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Titel: Development Genes and Evolution
Genre der Quelle: Zeitschrift
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Ort, Verlag, Ausgabe: Berlin : Springer-Verlag
Seiten: - Band / Heft: 225 (1) Artikelnummer: - Start- / Endseite: 55 - 62 Identifikator: ISSN: 0949-944X
CoNE: https://pure.mpg.de/cone/journals/resource/954926988121