The ATP13A2 Knockout HT29 Polyclonal Cells product comprises a CRISPR/Cas9-edited heterogeneous HT29 cell population with targeted disruption of the human ATP13A2 gene. This polyclonal pool, generated without single-cell cloning, provides a stable loss-of-function model for the lysosomal P5-type ATPase ATP13A2. It retains the parental line??s epithelial traits while enabling study of ATP13A2-dependent pathways in a colorectal adenocarcinoma background, avoiding clonal compensatory artifacts.
The HT29 parental cell line was derived from a colorectal adenocarcinoma of a 44-year-old Caucasian female. It displays epithelial morphology and intestinal marker expression, serving as a well-established model of intestinal epithelium. HT29 cells can differentiate into enterocyte-like phenotypes under confluent or butyrate-treated conditions, supporting polarized epithelial assays relevant to protein trafficking and barrier function.
ATP13A2, a lysosomal P5-ATPase, transports polyamines into lysosomes, regulated by TFEB, oxidative stress, and polyamine levels. It interacts with HDAC6, ??-synuclein, and LAMP2. Downstream, its loss impairs autophagic flux (LC3-II/p62 accumulation), lysosomal pH, cathepsin D activity, and promotes ??-synuclein aggregation, while disrupting Parkin/PINK1-mediated mitophagy and mitochondrial dynamics.
ATP13A2 knockout in HT29 disrupts polyamine transport and autophagy, recapitulating lysosomal dysfunction and ??-synuclein pathology seen in Parkinson??s disease and Kufor-Rakeb syndrome. This model enables dissection of lysosomal stress, protein aggregation, and mitochondrial dysfunction crosstalk in an epithelial system, complementing neuronal models and exploring peripheral contributions to neurodegeneration.
Applications include Parkinson??s disease modeling, autophagy research, and drug screening for lysosomal function enhancers. Assays such as Western blot (ATP13A2, ??-synuclein, LC3, p62), immunofluorescence (LAMP2, LC3 puncta), bafilomycin A1-based flux assays, polyamine transport measurements, MitoTracker/Seahorse mitochondrial analysis, and stress viability assays are compatible. For further assistance, contact Ascent Research.
