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dc.contributor.authorIttiprasert, W.
dc.contributor.authorMann, V.H.
dc.contributor.authorKarinshak, S.E.
dc.date.accessioned2019-03-29T14:47:38Z
dc.date.available2019-03-29T14:47:38Z
dc.date.issued2019
dc.identifier.urihttps://www.scopus.com/inward/record.uri?eid=2-s2.0-85060919508&doi=10.7554%2feLife.41337&partnerID=40&md5=e2dea6590a67e0f640bfdf5f64912d49
dc.identifier.urihttp://hdl.handle.net/10713/8718
dc.description.abstractCRISPR/Cas9-based genome editing has yet to be reported in species of the Platyhelminthes. We tested this approach by targeting omega-1 (ω1) of Schistosoma mansoni as proof of principle. This secreted ribonuclease is crucial for Th2 polarization and granuloma formation. Schistosome eggs were exposed to Cas9 complexed with guide RNA complementary to ω1 by electroporation or by transduction with lentiviral particles. Some eggs were also transfected with a single stranded donor template. Sequences of amplicons from gene-edited parasites exhibited Cas9-catalyzed mutations including homology directed repaired alleles, and other analyses revealed depletion of ω1 transcripts and the ribonuclease. Gene-edited eggs failed to polarize Th2 cytokine responses in macrophage/T-cell co-cultures, while the volume of pulmonary granulomas surrounding ω1-mutated eggs following tail-vein injection into mice was vastly reduced. Knock-out of ω1 and the diminished levels of these cytokines following exposure showcase the novel application of programmed gene editing for functional genomics in schistosomes.en_US
dc.description.sponsorshipNational Institute of Allergy and Infectious Diseases (R21AI109532) -Gabriel Rinaldi ; Thailand Research Fund (PHD/0011/2555) - Apisit Chaidee; Thailand Research Fund (PHD/0047/2556) - Pannathee Prangtaworn; Thailand Research Fund (PHD/00531/2556) - Rutchanee Rodpai; Wellcome (107475/Z/15/Z) - Karl F Hoffmann; Wellcome (WT 098051) - Matthew Berriman; MaxMind Inc - Paul J Brindley. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
dc.description.urihttps://dx.doi.org/10.7554/eLife.41337en_US
dc.language.isoen_USen_US
dc.publishereLifeen_US
dc.relation.ispartofeLife
dc.subjectCRISPR/Cas9en_US
dc.subjectgranulomaen_US
dc.subjectinfectious diseaseen_US
dc.subjectmicrobiologyen_US
dc.subjectomega-1en_US
dc.subjectribonucleaseen_US
dc.subjectSchistosoma mansonien_US
dc.subjectTh2-polarizationen_US
dc.titleProgrammed genome editing of the omega-1 ribonuclease of the blood fluke, Schistosoma mansonien_US
dc.typeArticleen_US
dc.identifier.doi10.7554/eLife.41337
dc.identifier.pmid30644357


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