Validation of a Miniaturized Permeability Assay Compatible with CRISPR-Mediated Genome-Wide Screen.

ABSTRACT: The impermeability of the luminal endothelial cell monolayer is crucial for the normal performance of the vascular and lymphatic systems. A key to this function is the integrity of the monolayer's intercellular junctions. The known repertoire of junction-regulating genes is incomplete. Current permeability assays are incompatible with high-throughput genome-wide screens that could identify these genes. To overcome these limitations, we designed a new permeability assay that consists of cell monolayers grown on ~150 μm microcarriers (MCs). Each MC functions as a miniature individual assay of permeability (MAP). We demonstrate that false-positive results can be minimized, and that MAP sensitivity to thrombin-induced increase in monolayer permeability is similar to the sensitivity of impedanc

SUBMITTER: Simonneau C 

PROVIDER: S-EPMC6775082 | biostudies-literature | 2019 Oct

REPOSITORIES: biostudies-literature