Transfected plasmids have reduced expression in cells deficient in SEPTIN 9 or ESCRT proteins
Transfection of cells with DNA plasmids typically involves the uptake of lipoparticles by endocytosis, followed by the inefficient escape of these particles from endosomes into the cytoplasm. We found that the expression of transfected plasmids was reduced in cells depleted of either SEPTIN 9 or proteins in the endosomal sorting complexes required for transport (ESCRT) pathway. The reduction in…
Transfected plasmids exhibit reduced expression in cells lacking SEPTIN 9 or ESCRT proteins, according to recent research. Unlike typical transfection processes, which rely on endocytosis and the inefficient release of lipoparticles from endosomes into the cytoplasm, cells deficient in SEPTIN 9 or ESCRT proteins show a significant decrease in plasmid expression.
This reduction cannot be attributed solely to impaired endocytosis. SEPTIN 9 depletion specifically diminishes the acidification of plasmid-containing compartments, implying that it primarily impacts the pH-sensitive escape of plasmids from endosomes. Additionally, the depletion of ESCRT proteins like VPS36 or ALIX leads to a marked reduction in transfected plasmid expression, coinciding with a decrease in colocalization between the plasmids and CHMP4, a crucial ESCRT protein involved in endosomal membrane remodeling during intraluminal vesicle formation.
Notably, the strong colocalization of transfected plasmid DNA with LC3B suggests that the default pathway for such material is autophagy.
Written by urgent.news from bioRxiv's reporting — not their text. Machine-written — may contain errors; check the original before relying on it.
