Programmed genome editing of the omega-1 ribonuclease of the blood fluke, Schistosoma mansoni
Abstract
CRISPR/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.Sponsors
National 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.Keyword
CRISPR/Cas9granuloma
infectious disease
microbiology
omega-1
ribonuclease
Schistosoma mansoni
Th2-polarization
Identifier to cite or link to this item
https://www.scopus.com/inward/record.uri?eid=2-s2.0-85060919508&doi=10.7554%2feLife.41337&partnerID=40&md5=e2dea6590a67e0f640bfdf5f64912d49; http://hdl.handle.net/10713/8718ae974a485f413a2113503eed53cd6c53
10.7554/eLife.41337
Scopus Count
Collections
Related articles
- CRISPR/Cas9-mediated genome editing of Schistosoma mansoni acetylcholinesterase.
- Authors: You H, Mayer JU, Johnston RL, Sivakumaran H, Ranasinghe S, Rivera V, Kondrashova O, Koufariotis LT, Du X, Driguez P, French JD, Waddell N, Duke MG, Ittiprasert W, Mann VH, Brindley PJ, Jones MK, McManus DP
- Issue date: 2021 Jan
- RNA-Guided AsCas12a- and SpCas9-Catalyzed Knockout and Homology Directed Repair of the Omega-1 Locus of the Human Blood Fluke, Schistosoma mansoni.
- Authors: Ittiprasert W, Chatupheeraphat C, Mann VH, Li W, Miller A, Ogunbayo T, Tran K, Alrefaei YN, Mentink-Kane M, Brindley PJ
- Issue date: 2022 Jan 6
- Molecular characterization of omega-1: a hepatotoxic ribonuclease from Schistosoma mansoni eggs.
- Authors: Fitzsimmons CM, Schramm G, Jones FM, Chalmers IW, Hoffmann KF, Grevelding CG, Wuhrer M, Hokke CH, Haas H, Doenhoff MJ, Dunne DW
- Issue date: 2005 Nov
- Analysis of cytokine mRNA expression during primary granuloma formation induced by eggs of Schistosoma mansoni.
- Authors: Wynn TA, Eltoum I, Cheever AW, Lewis FA, Gause WC, Sher A
- Issue date: 1993 Aug 1
- TALEN- and CRISPR-enhanced DNA homologous recombination for gene editing in zebrafish.
- Authors: Zhang Y, Huang H, Zhang B, Lin S
- Issue date: 2016