Protocol to induce de novo lipid droplets in Saccharomyces cerevisiae

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 conditi...

Descripción completa

Detalles Bibliográficos
Autores principales: Khatri, Amit, Choudhary, Vineet
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10687286/
https://www.ncbi.nlm.nih.gov/pubmed/37995191
http://dx.doi.org/10.1016/j.xpro.2023.102732
_version_ 1785151951560769536
author Khatri, Amit
Choudhary, Vineet
author_facet Khatri, Amit
Choudhary, Vineet
author_sort Khatri, Amit
collection PubMed
description 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.
format Online
Article
Text
id pubmed-10687286
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher Elsevier
record_format MEDLINE/PubMed
spelling pubmed-106872862023-11-30 Protocol to induce de novo lipid droplets in Saccharomyces cerevisiae Khatri, Amit Choudhary, Vineet STAR Protoc Protocol 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. Elsevier 2023-11-22 /pmc/articles/PMC10687286/ /pubmed/37995191 http://dx.doi.org/10.1016/j.xpro.2023.102732 Text en © 2023 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Khatri, Amit
Choudhary, Vineet
Protocol to induce de novo lipid droplets in Saccharomyces cerevisiae
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 Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10687286/
https://www.ncbi.nlm.nih.gov/pubmed/37995191
http://dx.doi.org/10.1016/j.xpro.2023.102732
work_keys_str_mv AT khatriamit protocoltoinducedenovolipiddropletsinsaccharomycescerevisiae
AT choudharyvineet protocoltoinducedenovolipiddropletsinsaccharomycescerevisiae

Ejemplares similares