The ACBD3 Knockout HT29 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal population of HT29 cells carrying a targeted disruption of the ACBD3 gene. This knock-out model abolishes expression of the Golgi scaffold protein ACBD3, enabling functional interrogation of its roles in Golgi architecture and intracellular trafficking. The polyclonal nature of the product provides a heterogeneous pool of edited alleles, representing a versatile tool for loss-of-function studies without the clonal selection bias associated with single-cell-derived lines. As a population-level reagent, it is well suited for biochemical assays, imaging-based phenotypic screens, and other applications where bulk cellular responses are of primary interest.
The parental HT29 cell line is a widely used human colorectal adenocarcinoma model derived from a primary tumor of the colon. These adherent epithelial cells display characteristic microvilli and retain wild-type p53, making them a relevant substrate for studying colorectal cancer biology and intestinal epithelial function. HT29 cells are capable of differentiation into enterocyte-like phenotypes under appropriate culture conditions, offering a platform to investigate differentiation-dependent processes. Their well-characterized signaling networks and robust growth properties facilitate reproducible experimental workflows in cancer research and drug development.
ACBD3 functions as a critical Golgi-resident scaffolding protein that directly interacts with phosphatidylinositol 4-kinase III?? (PI4KIII??), recruiting it to Golgi membranes to generate phosphatidylinositol-4-phosphate (PI4P). This lipid product is essential for maintaining Golgi structural integrity, regulating vesicle budding, and directing cargo sorting. Beyond PI4KIII??, ACBD3 forms complexes with giantin (GOLGB1), golgin-160 (GOLGA3), and RAB11, coordinating tethering and fusion events. The protein also engages with the Notch signaling modulator Numb and the amyloid precursor protein (APP), influencing proteolytic processing and signaling outputs. Viral proteins, including the Aichi virus 3A protein, hijack ACBD3 to redirect PI4P synthesis for replication organelle formation.
In the HT29 colorectal cancer background, loss of ACBD3 is predicted to impair Golgi organization and secretory trafficking, potentially altering the surface expression of growth factor receptors, adhesion molecules, and matrix metalloproteinases. Disruption of ACBD3-dependent PI4P pools may affect endosomal?CGolgi communication and downstream pathways such as Notch signaling, with consequences for proliferation, migration, and anchorage-independent growth. The interaction with APP suggests relevance to Alzheimer??s disease research, where altered Golgi processing can shift APP cleavage toward amyloidogenic fragments. This knockout model thus provides a unique system to dissect how Golgi dysfunction contributes to oncogenic phenotypes and neurodegenerative pathology in an epithelial context.
This product is suitable for a broad range of applications, including Golgi cell biology, cancer signaling, and host?Cvirus interaction studies. Typical assays include immunofluorescence staining of Golgi markers (giantin, GM130) to assess structural changes, Western blot confirmation of ACBD3 depletion, PI4P ELISA or lipidomic profiling, and cell viability or colony formation assays to evaluate tumorigenic potential. Researchers may also employ transwell migration assays, drug sensitivity testing (e.g., to chemotherapeutics or targeted agents), co-immunoprecipitation for interaction mapping, and transcriptomic analyses via RNA-seq or RT-qPCR. For additional details or technical support, please contact Ascent Research.
