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• W3099329529 abstract "Abstract Background Larval development in an intermediate host gastropod snail of the genus Biomphalaria is an obligatory component of the life cycle of Schistosoma mansoni . Understanding of the mechanism(s) of host defense may hasten the development of tools that block transmission of schistosomiasis. The allograft inflammatory factor 1, AIF, which is evolutionarily conserved and expressed in phagocytes, is a marker of macrophage activation in both mammals and invertebrates. AIF enhances cell proliferation and migration. The embryonic cell line, termed Bge, from Biomphalaria glabrata is a versatile resource for investigation of the snail-schistosome relationship since Bge exhibits a hemocyte-like phenotype. Hemocytes perform central roles in innate and cellular immunity in gastropods and in some cases can kill the parasite. However, the Bge cells do not kill the parasite in vitro . Methods Bge cells were transfected by electroporation with plasmid pCas -Bg AIFx4, encoding the Cas9 nuclease and a guide RNA specific for exon 4 of the B. glabrata AIF ( Bg AIF) gene. Transcript levels for Cas9 and for Bg AIF were monitored by quantitative reverse-transcription-PCR and, in parallel, adhesion of gene-edited Bge cells during co-culture with of schistosome sporocysts was assessed. Results Gene knockout manipulation induced gene-disrupting indels, frequently 1-2 bp insertions and/or 8-30 bp deletions, at the programmed target site; a range from 9 to 17% of the copies of the Bg AIF gene in the Bge population of cells were mutated. Transcript levels for Bg AIF were reduced by up to 73% (49.5±20.2% S.D, P ≤ 0.05, n =12). Adherence by Bg AIF gene-edited (Δ Bg AIF) Bge to sporocysts diminished in comparison to wild type cells, although cell morphology did not change. Specifically, as scored by a semi-quantitative cell adherence index (CAI), fewer Δ Bg AIF than control wild type cells adhered to sporocysts; control CAI, 2.66±0.10, Δ Bg AIF, 2.30±0.22 ( P ≤ 0.01). Conclusion The findings supported the hypothesis that Bg AIF plays a role in the adherence of B. glabrata hemocytes to sporocysts during schistosome infection in vitro . This demonstration of the activity of programmed gene editing will enable functional genomics approaches using CRISPR/Cas9 to investigate additional components of the snail-schistosome host-parasite relationship." @default.
• W3099329529 created "2020-11-23" @default.
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• W3099329529 date "2020-04-08" @default.
• W3099329529 modified "2023-10-18" @default.
• W3099329529 title "Diminished adherence of Biomphalaria glabrata embryonic cell line to sporocysts of Schistosoma mansoni following programmed knockout of the allograft inflammatory factor" @default.
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• W3099329529 doi "https://doi.org/10.1101/2020.04.07.029629" @default.
• W3099329529 hasPublicationYear "2020" @default.
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