Establishing a Cre/loxP-based genetic manipulation system for Acanthamoeba: Targeted genome editing and stable reporter expression.

ABSTRACT: Acanthamoeba is an opportunistic pathogen responsible for granulomatous amoebic encephalitis and amoebic keratitis. Despite its clinical significance, effective treatments remain challenging due to a limited understanding of its pathogenic mechanism. This study developed a genetic manipulation system in Acanthamoeba to facilitate gene function and drug screening studies. We applied the Cre/loxP system to integrate the gene encoding the tdTomato fluorescent protein into the genome of Acanthamoeba castellanii via homologous recombination. The polyubiquitin gene and its untranslated regions were identified and verified, after which the tdTomato gene was cloned between the untranslated regions of the polyubiquitin gene. The construct was then introduced into the Acanthamoeba genome using a mod

SUBMITTER: Aung JM 

PROVIDER: S-EPMC11895094 | biostudies-literature | 2025 Feb

REPOSITORIES: biostudies-literature