Gene editing in the nematode parasite Nippostrongylus brasiliensis using extracellular vesicles to deliver active Cas9/guide RNA complexes
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Published version
Author(s)
Hagen, Jana
Ghosh, Subhanita
Sarkies, Peter
Selkirk, Murray
Type
Journal Article
Abstract
Despite recent advances, animal-parasitic nematodes have thus far been largely refractory to genetic manipulation. We describe here a new approach providing proof of principle that CRISPR/Cas9-mediated gene editing of parasitic nematodes is achievable using vesicular stomatitis virus glycoprotein-pseudotyped extracellular vesicles for the delivery of Cas9-single guide ribonucleoprotein complexes. We demonstrate that extracellular vesicle-delivered ribonucleoproteins can be used to disrupt a secreted deoxyribonuclease in Nippostrogylus brasiliensis. Introduction of a repair template encoding multiple stop codons led to measurable reduction in expression of the targeted gene. Altered transcripts corresponding to the edited locus were detected by RT-PCR, demonstrating that vesicles can access cells of tissues actively expressing the gene of interest. These data provide evidence that this technique can be employed for targeted gene editing in N. brasiliensis, making this species genetically tractable for the first time, although further refinement will be necessary for routine and robust interrogation of gene function.
Date Issued
2023-01-26
Date Acceptance
2023-01-10
Citation
Frontiers in Parasitology, 2023, 2, pp.1-14
ISSN
2813-2424
Publisher
Frontiers
Start Page
1
End Page
14
Journal / Book Title
Frontiers in Parasitology
Volume
2
Copyright Statement
Copyright © 2023 Hagen, Ghosh, Sarkies and Selkirk. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
License URL
Identifier
https://www.frontiersin.org/articles/10.3389/fpara.2023.1071738/full
Publication Status
Published
Article Number
1071738
Date Publish Online
2023-01-26