Targeted mutagenesis in a human-parasitic nematode.
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 be...
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2017-10-01
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Series: | PLoS Pathogens |
Online Access: | https://doi.org/10.1371/journal.ppat.1006675 |
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doaj-437ca7f3760e4614ab2cb96bec8be7152021-04-21T17:54:40ZengPublic Library of Science (PLoS)PLoS Pathogens1553-73661553-73742017-10-011310e100667510.1371/journal.ppat.1006675Targeted mutagenesis in a human-parasitic nematode.Spencer S GangMichelle L CastellettoAstra S BryantEmily YangNicholas MancusoJacqueline B LopezMatteo PellegriniElissa A HallemParasitic 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.https://doi.org/10.1371/journal.ppat.1006675 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Spencer S Gang Michelle L Castelletto Astra S Bryant Emily Yang Nicholas Mancuso Jacqueline B Lopez Matteo Pellegrini Elissa A Hallem |
spellingShingle |
Spencer S Gang Michelle L Castelletto Astra S Bryant Emily Yang Nicholas Mancuso Jacqueline B Lopez Matteo Pellegrini Elissa A Hallem Targeted mutagenesis in a human-parasitic nematode. PLoS Pathogens |
author_facet |
Spencer S Gang Michelle L Castelletto Astra S Bryant Emily Yang Nicholas Mancuso Jacqueline B Lopez Matteo Pellegrini Elissa A Hallem |
author_sort |
Spencer S Gang |
title |
Targeted mutagenesis in a human-parasitic nematode. |
title_short |
Targeted mutagenesis in a human-parasitic nematode. |
title_full |
Targeted mutagenesis in a human-parasitic nematode. |
title_fullStr |
Targeted mutagenesis in a human-parasitic nematode. |
title_full_unstemmed |
Targeted mutagenesis in a human-parasitic nematode. |
title_sort |
targeted mutagenesis in a human-parasitic nematode. |
publisher |
Public Library of Science (PLoS) |
series |
PLoS Pathogens |
issn |
1553-7366 1553-7374 |
publishDate |
2017-10-01 |
description |
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. |
url |
https://doi.org/10.1371/journal.ppat.1006675 |
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