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CRISPR activation screen uncovers MARCKSL1 as a gauge for extracellular vesicle secretion

ABSTRACT: Extracellular vesicles (EVs) are crucial mediators of cell-cell communication that originate through one of two pathways: outward budding from the plasma membrane (PM) to yield microvesicles, or fusion of mature (late) endosomes with the PM for exosome release. How cells balance these two membrane pathways to optimize their EV repertoire remains unclear. To explore this, we performed a genome-wide CRISPR activation screen for genes that increase cell surface levels of the tetraspanin CD63—an EV marker that shuttles between late endosomes and the PM. Using this approach, we identified a membrane adaptor protein MARCKSL1 as a positive regulator of CD63 cell surface display. Follow-up ablation and overexpression studies demonstrated that MARCKSL1 potentiates EV secretion from the PM, in part at the expense of late endosome fusion with the PM. Mechanistically, we demonstrate that MARCKLS1 exploits its PM-binding effector domain to upregulate cell surface expression of CD63 and suppress late endosome—PM fusion. Furthermore, using proximity-based biotinylation proteomics, we identified the PM bridging protein Radixin and the SNARE-associated fusion component STXBP3 as key collaborators of MARCKSL1 in the context of PM homeostasis. Collectively, our findings reveal new mechanistic underpinnings of PM remodeling and implicate MARCKSL1 as a molecular gauge between different platforms for EV biogenesis.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: Rayman Tjokrodirijo

LAB HEAD: Peter A. van Veelen

PROVIDER: PXD066258 | Pride | 2026-06-18

REPOSITORIES: Pride

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			Action	DRS
	20250514_104833_P1919_Extracelular_vesicles.sne	Other
	A20252001840a.raw	Raw
	A20252001842a.raw	Raw
	A20252001844a.raw	Raw
	A20252001846a.raw	Raw

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