Title |
Gene inactivation using the CRISPR/Cas9 system in the nematode Pristionchus pacificus
|
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Published in |
Development Genes and Evolution, December 2014
|
DOI | 10.1007/s00427-014-0486-8 |
Pubmed ID | |
Authors |
Hanh Witte, Eduardo Moreno, Christian Rödelsperger, Jungeun Kim, Jin-Soo Kim, Adrian Streit, Ralf J. Sommer |
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. |
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Geographical breakdown
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United Kingdom | 2 | 40% |
Unknown | 3 | 60% |
Demographic breakdown
Type | Count | As % |
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Members of the public | 3 | 60% |
Science communicators (journalists, bloggers, editors) | 2 | 40% |
Mendeley readers
Geographical breakdown
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United States | 1 | 1% |
Australia | 1 | 1% |
Unknown | 85 | 98% |
Demographic breakdown
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Student > Ph. D. Student | 23 | 26% |
Researcher | 13 | 15% |
Student > Bachelor | 10 | 11% |
Student > Master | 8 | 9% |
Student > Doctoral Student | 4 | 5% |
Other | 9 | 10% |
Unknown | 20 | 23% |
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Biochemistry, Genetics and Molecular Biology | 19 | 22% |
Immunology and Microbiology | 3 | 3% |
Neuroscience | 3 | 3% |
Chemistry | 2 | 2% |
Other | 1 | 1% |
Unknown | 19 | 22% |