{"doi":"10.1371/journal.ppat.1010013","title":"Novel regulators of PrPC biosynthesis revealed by genome-wide RNA interference","abstract":"<jats:p>The cellular prion protein PrP<jats:sup>C</jats:sup> is necessary for prion replication, and its reduction greatly increases life expectancy in animal models of prion infection. Hence the factors controlling the levels of PrP<jats:sup>C</jats:sup> may represent therapeutic targets against human prion diseases. Here we performed an arrayed whole-transcriptome RNA interference screen to identify modulators of PrP<jats:sup>C</jats:sup> expression. We cultured human U251-MG glioblastoma cells in the presence of 64’752 unique siRNAs targeting 21’584 annotated human genes, and measured PrP<jats:sup>C</jats:sup> using a one-pot fluorescence-resonance energy transfer immunoassay in 51’128 individual microplate wells. This screen yielded 743 candidate regulators of PrP<jats:sup>C</jats:sup>. When downregulated, 563 of these candidates reduced and 180 enhanced PrP<jats:sup>C</jats:sup> expression. Recursive candidate attrition through multiple secondary screens yielded 54 novel regulators of PrP<jats:sup>C</jats:sup>, 9 of which were confirmed by CRISPR interference as robust regulators of PrP<jats:sup>C</jats:sup> biosynthesis and degradation. The phenotypes of 6 of the 9 candidates were inverted in response to transcriptional activation using CRISPRa. The RNA-binding post-transcriptional repressor Pumilio-1 was identified as a potent limiter of PrP<jats:sup>C</jats:sup> expression through the degradation of <jats:italic>PRNP</jats:italic> mRNA. Because of its hypothesis-free design, this comprehensive genetic-perturbation screen delivers an unbiased landscape of the genes regulating PrP<jats:sup>C</jats:sup> levels in cells, most of which were unanticipated, and some of which may be amenable to pharmacological targeting in the context of antiprion therapies.</jats:p>","journal":"PLOS Pathogens","year":2021,"id":642354,"datarank":0.38474240361923057,"base_score":2.5649493574615367,"endowment":2.5649493574615367,"self_citation_contribution":0.38474240361923057,"citation_network_contribution":0.0,"self_endowment_contribution":0.38474240361923057,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":12,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":688269,"name":"Merve Avar","orcid":"0000-0003-4665-5558","position":1,"is_corresponding":false},{"id":737052,"name":"Daniel Pease","orcid":"0000-0001-6632-7063","position":2,"is_corresponding":false},{"id":1670720,"name":"Ashutosh Dhingra","orcid":null,"position":3,"is_corresponding":false},{"id":1247246,"name":"Jiang‐An Yin","orcid":"0000-0002-2134-8693","position":4,"is_corresponding":false},{"id":1670721,"name":"Elke Schaper","orcid":"0000-0003-3734-2496","position":5,"is_corresponding":false},{"id":1670722,"name":"Berre Doğançay","orcid":null,"position":6,"is_corresponding":false},{"id":737045,"name":"Marc Emmenegger","orcid":"0000-0002-6073-8811","position":7,"is_corresponding":false},{"id":1670723,"name":"Anna Spinelli","orcid":"0000-0002-8829-7405","position":8,"is_corresponding":false},{"id":1670724,"name":"Kevin Maggi","orcid":null,"position":9,"is_corresponding":false},{"id":737051,"name":"Andra Chincisan","orcid":"0000-0003-3170-6500","position":10,"is_corresponding":false},{"id":300804,"name":"Simon Mead","orcid":"0000-0002-4326-1468","position":11,"is_corresponding":false},{"id":688270,"name":"Simone Hornemann","orcid":"0000-0002-2674-9891","position":12,"is_corresponding":false},{"id":40733,"name":"Peter Heutink","orcid":"0000-0001-5218-1737","position":13,"is_corresponding":false},{"id":300784,"name":"Adriano Aguzzi","orcid":"0000-0002-0344-6708","position":14,"is_corresponding":false},{"id":437381,"name":"Daniel Heinzer","orcid":"0000-0002-3282-4042","position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Novel regulators of PrPC biosynthesis revealed by genome-wide RNA interference","abstract":"<jats:p>The cellular prion protein PrP<jats:sup>C</jats:sup> is necessary for prion replication, and its reduction greatly increases life expectancy in animal models of prion infection. Hence the factors controlling the levels of PrP<jats:sup>C</jats:sup> may represent therapeutic targets against human prion diseases. Here we performed an arrayed whole-transcriptome RNA interference screen to identify modulators of PrP<jats:sup>C</jats:sup> expression. We cultured human U251-MG glioblastoma cells in the presence of 64’752 unique siRNAs targeting 21’584 annotated human genes, and measured PrP<jats:sup>C</jats:sup> using a one-pot fluorescence-resonance energy transfer immunoassay in 51’128 individual microplate wells. This screen yielded 743 candidate regulators of PrP<jats:sup>C</jats:sup>. When downregulated, 563 of these candidates reduced and 180 enhanced PrP<jats:sup>C</jats:sup> expression. Recursive candidate attrition through multiple secondary screens yielded 54 novel regulators of PrP<jats:sup>C</jats:sup>, 9 of which were confirmed by CRISPR interference as robust regulators of PrP<jats:sup>C</jats:sup> biosynthesis and degradation. 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