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CRISPR screens for lipid regulators reveal a role for ER-bound SNX13 in lysosomal cholesterol export.

Albert LuFrank HsiehBikal R SharmaSydney R VaughnCarles RenteroSuzanne R Pfeffer
Published in: The Journal of cell biology (2021)
We report here two genome-wide CRISPR screens performed to identify genes that, when knocked out, alter levels of lysosomal cholesterol or bis(monoacylglycero)phosphate. In addition, these screens were also performed under conditions of NPC1 inhibition to identify modifiers of NPC1 function in lysosomal cholesterol export. The screens confirm tight coregulation of cholesterol and bis(monoacylglycero)phosphate in cells and reveal an unexpected role for the ER-localized SNX13 protein as a negative regulator of lysosomal cholesterol export and contributor to ER-lysosome membrane contact sites. In the absence of NPC1 function, SNX13 knockdown redistributes lysosomal cholesterol and is accompanied by triacylglycerol-rich lipid droplet accumulation and increased lysosomal bis(monoacylglycero)phosphate. These experiments provide unexpected insight into the regulation of lysosomal lipids and modification of these processes by novel gene products.
Keyphrases
  • genome wide
  • dna methylation
  • low density lipoprotein
  • copy number
  • high throughput
  • ionic liquid
  • transcription factor
  • endoplasmic reticulum
  • cell death
  • oxidative stress
  • cell cycle arrest