Mating-based Overexpression Library Screening in Yeast
| Autor: | Abagail Chumley, Shulin Ju, Elliott Hayden, Quan Zhong, Shuzhen Chen |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
General Chemical Engineering Computational biology Saccharomyces cerevisiae Biology General Biochemistry Genetics and Molecular Biology 03 medical and health sciences Plasmid medicine Genetics Humans In patient Proteostasis Deficiencies Cytotoxicity Gene Gene Library General Immunology and Microbiology General Neuroscience Neurodegeneration Proteins Neurodegenerative Diseases medicine.disease Budding yeast Yeast 030104 developmental biology Protein folding |
| Zdroj: | J Vis Exp |
| ISSN: | 1940-087X |
| Popis: | Budding yeast has been widely used as a model in studying proteins associated with human diseases. Genome-wide genetic screening is a powerful tool commonly used in yeast studies. The expression of a number of neurodegenerative disease-associated proteins in yeast causes cytotoxicity and aggregate formation, recapitulating findings seen in patients with these disorders. Here, we describe a method for screening a yeast model of the Amyotrophic Lateral Sclerosis-associated protein FUS for modifiers of its toxicity. Instead of using transformation, this new screening platform relies on the mating of yeast to introduce an arrayed library of plasmids into the yeast model. The mating method has two clear advantages: first, it is highly efficient; second, the pre-transformed arrayed library of plasmids can be stored for long-term as a glycerol stock, and quickly applied to other screens without the labor-intensive step of transformation into the yeast model each time. We demonstrate how this method can successfully be used to screen for genes that modify the toxicity of FUS. |
| Databáze: | OpenAIRE |
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