CRISPR knockout screen implicates three genes in lysosome function.

ABSTRACT: Defective biosynthesis of the phospholipid PI(3,5)P₂ underlies neurological disorders characterized by cytoplasmic accumulation of large lysosome-derived vacuoles. To identify novel genetic causes of lysosomal vacuolization, we developed an assay for enlargement of the lysosome compartment that is amenable to cell sorting and pooled screens. We first demonstrated that the enlarged vacuoles that accumulate in fibroblasts lacking FIG4, a PI(3,5)P₂ biosynthetic factor, have a hyperacidic pH compared to normal cells'. We then carried out a genome-wide knockout screen in human HAP1 cells for accumulation of acidic vesicles by FACS sorting. A pilot screen captured fifteen genes, including VAC14, a previously identified cause of endolysosomal vacuolization. Three genes not p

SUBMITTER: Lenk GM 

PROVIDER: S-EPMC6610096 | biostudies-literature | 2019 Jul

REPOSITORIES: biostudies-literature