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PLoS Pathog. 2017 Oct 10;13(10):e1006675. doi: 10.1371/journal.ppat.1006675. eCollection 2017 Oct.

Targeted mutagenesis in a human-parasitic nematode.

Author information

1
Molecular Biology Institute, University of California, Los Angeles, California, United States of America.
2
Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, California, United States of America.
3
Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California, Los Angeles, California, United States of America.
4
Department of Molecular, Cell and Developmental Biology, University of California, Los Angeles, California, United States of America.

Abstract

Parasitic nematodes infect over 1 billion people worldwide and cause some of the most common neglected tropical diseases. Despite their prevalence, our understanding of the biology of parasitic nematodes has been limited by the lack of tools for genetic intervention. In particular, it has not yet been possible to generate targeted gene disruptions and mutant phenotypes in any parasitic nematode. Here, we report the development of a method for introducing CRISPR-Cas9-mediated gene disruptions in the human-parasitic threadworm Strongyloides stercoralis. We disrupted the S. stercoralis twitchin gene unc-22, resulting in nematodes with severe motility defects. Ss-unc-22 mutations were resolved by homology-directed repair when a repair template was provided. Omission of a repair template resulted in deletions at the target locus. Ss-unc-22 mutations were heritable; we passed Ss-unc-22 mutants through a host and successfully recovered mutant progeny. Using a similar approach, we also disrupted the unc-22 gene of the rat-parasitic nematode Strongyloides ratti. Our results demonstrate the applicability of CRISPR-Cas9 to parasitic nematodes, and thereby enable future studies of gene function in these medically relevant but previously genetically intractable parasites.

PMID:
29016680
PMCID:
PMC5650185
DOI:
10.1371/journal.ppat.1006675
[Indexed for MEDLINE]
Free PMC Article

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