2020
DOI: 10.26508/lsa.202000814
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Prion infection, transmission, and cytopathology modeled in a low-biohazard human cell line

Abstract: Transmission of prion infectivity to susceptible murine cell lines has simplified prion titration assays and has greatly reduced the need for animal experimentation. However, murine cell models suffer from technical and biological constraints. Human cell lines might be more useful, but they are much more biohazardous and are often poorly infectible. Here, we describe the human clonal cell line hovS, which lacks the human PRNP gene and expresses instead the ovine PRNP VRQ allele. HovS cells were highly suscepti… Show more

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Cited by 12 publications
(10 citation statements)
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“…The proof that PrP Sc represents infectivity was first based on biomathematical and extensive biochemical work with hamster scrapie [ 1 5 ]. Definitive proof that the presence of PrP C is a prerequisite for TSE infection was presented from PrP-less mice, goats and cells, and by the production of infectivity from recombinantly expressed and purified PrP [ 6 14 ]. Another argument for the validity of the role of PrP in the agent is the close relation between susceptibility/resistance in e.g.…”
Section: Introductionmentioning
confidence: 99%
“…The proof that PrP Sc represents infectivity was first based on biomathematical and extensive biochemical work with hamster scrapie [ 1 5 ]. Definitive proof that the presence of PrP C is a prerequisite for TSE infection was presented from PrP-less mice, goats and cells, and by the production of infectivity from recombinantly expressed and purified PrP [ 6 14 ]. Another argument for the validity of the role of PrP in the agent is the close relation between susceptibility/resistance in e.g.…”
Section: Introductionmentioning
confidence: 99%
“…For instance, rabbit RK13 cells become susceptible to chronic wasting disease and scrapie prions upon expression of elk or sheep PrP C , respectively ( 29 , 30 , 31 ). Similar paradigms have been developed using cells derived from PrP knockout mice or gene-edited PrP C -null (PrP −/− ) cell lines as a starting point ( 32 , 33 , 34 , 35 , 36 ). In each of these cases, selection of stably transfected cells expressing the desired PrP C is typically accomplished by coexpression of an antibiotic resistance gene and subsequent antibiotic treatment.…”
mentioning
confidence: 99%
“…For infection of cells with different strains of prions, a previously established protocol was followed (Avar et al, 2020). Cells were seeded in 6-well plates and the infection occurred for all prion strains (22L, RML, PG127) at the same weight/volume ratio of 0.25% brain homogenate containing prions or non-infectious brain homogenate (NBH) in a total culture media volume of 1.5 mL.…”
Section: Methodsmentioning
confidence: 99%