Protocol to induce de novo lipid droplets in Saccharomyces cerevisiae
Summary: The mechanisms underlying lipid droplet (LD) biogenesis in the endoplasmic reticulum are not completely understood. Here, we present a protocol for inducing de novo LDs containing either triacylglycerol (TAG) or sterol esters (SE) in Saccharomyces cerevisiae. We describe steps for generatin...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-12-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166723006998 |
| _version_ | 1797449391803465728 |
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| author | Amit Khatri Vineet Choudhary |
| author_facet | Amit Khatri Vineet Choudhary |
| author_sort | Amit Khatri |
| collection | DOAJ |
| description | Summary: The mechanisms underlying lipid droplet (LD) biogenesis in the endoplasmic reticulum are not completely understood. Here, we present a protocol for inducing de novo LDs containing either triacylglycerol (TAG) or sterol esters (SE) in Saccharomyces cerevisiae. We describe steps for generating conditional yeast mutants by performing gene knockout and introducing galactose-inducible promoter to produce TAG- or SE-containing LDs. We detail the strategy to generate fluorescent LD marker protein to visualize newly formed droplets by fluorescence microscopy. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-03-09T14:24:24Z |
| format | Article |
| id | doaj.art-00078838020842a99ed117ffdcb9c144 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-03-09T14:24:24Z |
| publishDate | 2023-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-00078838020842a99ed117ffdcb9c1442023-11-28T07:27:02ZengElsevierSTAR Protocols2666-16672023-12-0144102732Protocol to induce de novo lipid droplets in Saccharomyces cerevisiaeAmit Khatri0Vineet Choudhary1Lipid Metabolism Laboratory, Department of Biotechnology, All India Institute of Medical Sciences (AIIMS), New Delhi 110029, IndiaLipid Metabolism Laboratory, Department of Biotechnology, All India Institute of Medical Sciences (AIIMS), New Delhi 110029, India; Corresponding authorSummary: The mechanisms underlying lipid droplet (LD) biogenesis in the endoplasmic reticulum are not completely understood. Here, we present a protocol for inducing de novo LDs containing either triacylglycerol (TAG) or sterol esters (SE) in Saccharomyces cerevisiae. We describe steps for generating conditional yeast mutants by performing gene knockout and introducing galactose-inducible promoter to produce TAG- or SE-containing LDs. We detail the strategy to generate fluorescent LD marker protein to visualize newly formed droplets by fluorescence microscopy. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166723006998cell Biologymodel OrganismsMolecular Biologyprotein Biochemistry |
| spellingShingle | Amit Khatri Vineet Choudhary Protocol to induce de novo lipid droplets in Saccharomyces cerevisiae STAR Protocols cell Biology model Organisms Molecular Biology protein Biochemistry |
| title | Protocol to induce de novo lipid droplets in Saccharomyces cerevisiae |
| title_full | Protocol to induce de novo lipid droplets in Saccharomyces cerevisiae |
| title_fullStr | Protocol to induce de novo lipid droplets in Saccharomyces cerevisiae |
| title_full_unstemmed | Protocol to induce de novo lipid droplets in Saccharomyces cerevisiae |
| title_short | Protocol to induce de novo lipid droplets in Saccharomyces cerevisiae |
| title_sort | protocol to induce de novo lipid droplets in saccharomyces cerevisiae |
| topic | cell Biology model Organisms Molecular Biology protein Biochemistry |
| url | http://www.sciencedirect.com/science/article/pii/S2666166723006998 |
| work_keys_str_mv | AT amitkhatri protocoltoinducedenovolipiddropletsinsaccharomycescerevisiae AT vineetchoudhary protocoltoinducedenovolipiddropletsinsaccharomycescerevisiae |