Programmed genome editing of the omega-1 ribonuclease of the blood fluke, Schistosoma mansoni .

Autor:	Ittiprasert W; Department of Microbiology, Immunology and Tropical Medicine, School of Medicine and Health Sciences, George Washington University, Washington, DC, United States.; Research Center for Neglected Diseases of Poverty, School of Medicine and Health Sciences, George Washington University, Washington, DC, United States., Mann VH; Department of Microbiology, Immunology and Tropical Medicine, School of Medicine and Health Sciences, George Washington University, Washington, DC, United States.; Research Center for Neglected Diseases of Poverty, School of Medicine and Health Sciences, George Washington University, Washington, DC, United States., Karinshak SE; Research Center for Neglected Diseases of Poverty, School of Medicine and Health Sciences, George Washington University, Washington, DC, United States., Coghlan A; Wellcome Sanger Institute, Wellcome Genome Campus, Hinxton, United Kingdom., Rinaldi G; Wellcome Sanger Institute, Wellcome Genome Campus, Hinxton, United Kingdom., Sankaranarayanan G; Wellcome Sanger Institute, Wellcome Genome Campus, Hinxton, United Kingdom., Chaidee A; Research Center for Neglected Diseases of Poverty, School of Medicine and Health Sciences, George Washington University, Washington, DC, United States.; Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand., Tanno T; Department of Surgery, University of Maryland, Baltimore, United States.; Institute of Human Virology, University of Maryland, Baltimore, United States., Kumkhaek C; Cellular and Molecular Therapeutics Laboratory, National Heart, Lungs and Blood Institute, National Institutes of Health, Bethesda, United States., Prangtaworn P; Research Center for Neglected Diseases of Poverty, School of Medicine and Health Sciences, George Washington University, Washington, DC, United States.; Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand., Mentink-Kane MM; Schistosomiasis Resource Center, Biomedical Research Institute, Rockville, United States., Cochran CJ; Department of Microbiology, Immunology and Tropical Medicine, School of Medicine and Health Sciences, George Washington University, Washington, DC, United States., Driguez P; Wellcome Sanger Institute, Wellcome Genome Campus, Hinxton, United Kingdom., Holroyd N; Wellcome Sanger Institute, Wellcome Genome Campus, Hinxton, United Kingdom., Tracey A; Wellcome Sanger Institute, Wellcome Genome Campus, Hinxton, United Kingdom., Rodpai R; Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand., Everts B; Department of Parasitology, Leiden University Medical Center, Leiden, Netherlands., Hokke CH; Department of Parasitology, Leiden University Medical Center, Leiden, Netherlands., Hoffmann KF; Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, Aberystwyth, United Kingdom., Berriman M; Wellcome Sanger Institute, Wellcome Genome Campus, Hinxton, United Kingdom., Brindley PJ; Department of Microbiology, Immunology and Tropical Medicine, School of Medicine and Health Sciences, George Washington University, Washington, DC, United States.
Jazyk:	angličtina
Zdroj:	ELife [Elife] 2019 Jan 15; Vol. 8. Date of Electronic Publication: 2019 Jan 15.
DOI:	10.7554/eLife.41337
Abstrakt:	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. Competing Interests: WI, VM, SK, AC, GR, GS, AC, TT, CK, PP, MM, CC, PD, NH, AT, RR, BE, CH, KH, MB, PB No competing interests declared
Databáze:	MEDLINE
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