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@ARTICLE{Heinzer:162908,
author = {Heinzer, Daniel and Avar, Merve and Pease, Daniel Patrick
and Dhingra, Ashutosh and Yin, Jiang-An and Schaper, Elke
and Doğançay, Berre and Emmenegger, Marc and Spinelli,
Anna and Maggi, Kevin and Chincisan, Andra and Mead, Simon
and Hornemann, Simone and Heutink, Peter and Aguzzi,
Adriano},
title = {{N}ovel regulators of {P}r{PC} biosynthesis revealed by
genome-wide {RNA} interference.},
journal = {PLoS pathogens},
volume = {17},
number = {10},
issn = {1553-7374},
address = {Lawrence, Kan.},
publisher = {PLoS},
reportid = {DZNE-2021-01563},
pages = {e1010013},
year = {2021},
note = {(CC BY)},
abstract = {The cellular prion protein PrPC 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 PrPC may represent
therapeutic targets against human prion diseases. Here we
performed an arrayed whole-transcriptome RNA interference
screen to identify modulators of PrPC expression. We
cultured human U251-MG glioblastoma cells in the presence of
64'752 unique siRNAs targeting 21'584 annotated human genes,
and measured PrPC using a one-pot fluorescence-resonance
energy transfer immunoassay in 51'128 individual microplate
wells. This screen yielded 743 candidate regulators of PrPC.
When downregulated, 563 of these candidates reduced and 180
enhanced PrPC expression. Recursive candidate attrition
through multiple secondary screens yielded 54 novel
regulators of PrPC, 9 of which were confirmed by CRISPR
interference as robust regulators of PrPC 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 PrPC
expression through the degradation of PRNP mRNA. Because of
its hypothesis-free design, this comprehensive
genetic-perturbation screen delivers an unbiased landscape
of the genes regulating PrPC levels in cells, most of which
were unanticipated, and some of which may be amenable to
pharmacological targeting in the context of antiprion
therapies.},
keywords = {Cell Line / Gene Expression Regulation: physiology /
Genome-Wide Association Study / Humans / PrPC Proteins:
biosynthesis / RNA Interference / RNA-Binding Proteins:
metabolism},
cin = {AG Heutink 1},
ddc = {610},
cid = {I:(DE-2719)1210002},
pnm = {354 - Disease Prevention and Healthy Aging (POF4-354)},
pid = {G:(DE-HGF)POF4-354},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:34705895},
pmc = {pmc:PMC8575309},
doi = {10.1371/journal.ppat.1010013},
url = {https://pub.dzne.de/record/162908},
}