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• W2917711557 abstract "Identification and degradation of misfolded proteins by the ubiquitin-proteasome system (UPS) is crucial for maintaining proteostasis, but only a handful of UPS components have been linked to the recognition of specific substrates. Studies in Saccharomyces cerevisiae using systematic perturbation of nonessential genes have uncovered UPS components that recognize and ubiquitylate model substrates of the UPS; however, similar analyses in metazoans have been limited. In this chapter, we describe methods for using CRISPR/Cas9 technology combined with genome-wide high complexity single guide (sgRNA) libraries and a transcriptional shutoff strategy for phenotypic selection based on kinetic measurements of protein turnover to identify the genes required to degrade model clients of the mammalian ER-associated degradation system. We also discuss considerations for screen design, execution, and interpretation." @default.
• W2917711557 created "2019-03-02" @default.
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• W2917711557 date "2019-01-01" @default.
• W2917711557 modified "2023-10-14" @default.
• W2917711557 title "Methods for genetic analysis of mammalian ER-associated degradation" @default.
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• W2917711557 doi "https://doi.org/10.1016/bs.mie.2019.01.006" @default.
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