Identification of Proteins Involved in Cell Membrane Permeabilization by Nanosecond Electric Pulses (nsEP)

• DOI: 10.3390/ijms24119191
• Publication Date: 2023-05-25T06:58:48Z
• Authors: Giedre Silkuniene; Uma M. Mangalanathan; Alessandra Rossi; Peter A. Mollica; Andrei G. Pakhomov; Olga Pakhomova
• Keywords: Cell membrane; Nuclease

The study was aimed at identifying endogenous proteins which assist or impede the permeabilized state in cell membrane disrupted by nsEP (20 40 pulses, 300 ns width, 7 kV/cm). We employed a LentiArray CRISPR library to generate knockouts (KOs) of 316 genes encoding for U937 human monocytes stably expressing Cas9 nuclease. extent permeabilization measured uptake Yo-Pro-1 (YP) dye and compared sham-exposed KOs control cells transduced with non-targeting (scrambled) gRNA. Only two KOs, SCNN1A CLCA1 genes, showed statistically significant reduction YP uptake. respective could be part electropermeabilization lesions increase their lifespan. In contrast, as many 39 were identified likely hits increased uptake, meaning that contributed stability repair after nsEP. expression level eight different types strong correlation (R > 0.9, p < 0.02) LD50 lethal treatments, potentially used criterion selectivity efficiency hyperplasia ablations