Development of Approaches for Transgene Expression in the Pathogenic Free-Living Amoeba Naegleria fowleri.

ABSTRACT: The absence of molecular tools for manipulation of gene expression in the pathogenic free-living amoeba Naegleria fowleri has historically limited our understanding of gene function in the organism and has coincidently impacted the identification of potential druggable pathways and proteins. Here, we describe the development of approaches for the generation of transgenic amoebae using polyethyleneimine nanoparticles to deliver plasmids designed to confer antibiotic resistance and fluorescence to the cells. Through a series of optimization steps, we found that transfection of plasmids encoding the fluorescent protein mCherry fused by a T2A self-cleaving peptide to a codon-optimized puromycin acetyltransferase selectable marker yielded fluorescent cells that were resistant up to 100 µ

SUBMITTER: Palmentiero CM 

PROVIDER: S-EPMC12845041 | biostudies-literature | 2025 Dec

REPOSITORIES: biostudies-literature