GOPC Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the Golgi-associated PDZ and coiled-coil motif-containing protein (GOPC, also known as CAL) in the HT29 human colon adenocarcinoma cell line. This loss-of-function model enables the investigation of GOPC-dependent pathways without introducing monoclonal selection bias, preserving the genetic heterogeneity inherent to polyclonal populations. The CRISPR/Cas9-mediated gene disruption provides a robust tool for studying GOPC’s roles in vesicular trafficking, cell polarity, and protein degradation mechanisms.
The HT29 cell line is a well-differentiated, epithelial model derived from a primary colorectal adenocarcinoma of a 44-year-old female. These cells retain characteristics of intestinal epithelial cells, including the capacity for mucus production and the formation of polarized monolayers with functional tight junctions. As a tumorigenic cell line, HT29 is widely used to study colorectal cancer biology, intestinal barrier function, and the molecular mechanisms underlying epithelial cell polarity and protein trafficking.
GOPC (CAL) functions as a Golgi-associated scaffold protein that critically regulates intracellular trafficking and cell polarity. It facilitates the lysosomal degradation of the cystic fibrosis transmembrane conductance regulator (CFTR) by interacting with syntaxin 6 (STX6) and the Golgi reassembly stacking protein 1 (GORASP1), thereby controlling CFTR surface expression. Additionally, GOPC acts as a scaffold for the Par polarity complex, interacting with PARD3, PARD6A, and atypical protein kinase C (PRKCZ/??), which are essential for tight junction assembly and apical-basal polarity. Upstream signals from Golgi stress and cellular trafficking demands modulate GOPC activity, while downstream effects include the regulation of CFTR trafficking, vesicle tethering, and Golgi structural integrity. Key pathway components include CDC42, a small GTPase that activates the Par complex, further linking GOPC to actin cytoskeletal dynamics and polarity signaling.
In the HT29 intestinal epithelial context, loss of GOPC disrupts CFTR trafficking, potentially altering chloride secretion and mucus homeostasis, which are relevant to cystic fibrosis pathophysiology. Moreover, because HT29 cells form polarized monolayers, GOPC knockout perturbs the Par complex-mediated tight junction assembly, compromising epithelial barrier integrity and cell polarity. This dual impact makes the model particularly valuable for dissecting the molecular crosstalk between protein trafficking, polarity establishment, and tumorigenic properties in colorectal cancer.
Typical research applications include western blotting and immunofluorescence analysis of CFTR and tight junction proteins to assess trafficking and localization defects, co-immunoprecipitation studies to map GOPC interactor networks, and barrier integrity assays such as transepithelial electrical resistance (TEER) measurements. Additionally, migration and invasion assays can explore the role of GOPC in metastatic behavior. This polyclonal knockout cell population thus provides a versatile platform for mechanistic studies in cell biology and drug discovery. For technical specifications or ordering information, please contact Ascent Research.
