Mammalian cell culture and transfection for stable cell lines generation v1

Autosomal recessive mutations in PTEN-induced kinase 1 (PINK1) are linked to early-onset Parkinson's disease (PD) [1]. Upon mitochondrial depolarization, PINK1 activates through autophosphorylation and stabilization on mitochonria [2]. Pink1 phosphorylates ubiquitin and Parkin, triggering mitophagy to remove damaged mitochondria in PD [3]. To delve deeper into the impact of PINK1 mutations, a PINK1 knockout (KO) HeLa cell line was utilized as a model system. Additionally, stable cell lines with mutated PINK1 were established to explore differences in functional activity and the formation of the PINK1-TOM complex between wild-type PINK1 and its mutant variants.