Journal
PLOS PATHOGENS
Volume 13, Issue 10, Pages -Publisher
PUBLIC LIBRARY SCIENCE
DOI: 10.1371/journal.ppat.1006675
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Funding
- Whitcome Predoctoral Training Program
- UCLA Molecular Biology Institute
- Ruth L. Kirschstein National Research Service Award [AI007323]
- National Science Foundation East Asia and Pacific Summer Institute Fellowship [1414655]
- UCLA Undergraduate Research Scholar Program
- UCLA-Howard Hughes Medical Institute Pathways
- Burroughs-Wellcome Fund Investigators in the Pathogenesis of Disease Award, NIH New Innovator [1DP2DC014596]
- Howard Hughes Medical Institute Faculty Scholar Award
- Office Of Internatl Science &Engineering
- Office Of The Director [1414655] Funding Source: National Science Foundation
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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.
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