The GORASP1 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population with disruption of the GORASP1 gene, providing a loss-of-function model in HT29 human colorectal adenocarcinoma cells. This polyclonal pool is generated by CRISPR/Cas9-mediated gene targeting, yielding a heterogeneous population with GORASP1 knockout, enabling study of gene function without clonal selection. As a polyclonal knockout reagent, it offers a robust tool for investigating Golgi biology, protein trafficking, and cancer cell behavior in a physiologically relevant epithelial context.
The host cell line HT29 is a well-characterized human colorectal adenocarcinoma epithelial cell line originally established from a primary tumor of a 44-year-old female. These cells serve as a classic intestinal epithelial model and are extensively used in colorectal cancer research, mucin secretion studies, and epithelial barrier function assays. Renowned for their polarized morphology and ability to form tight junctions, HT29 cells are a preferred model for analyzing tumor cell migration, invasion, and drug sensitivity, making them particularly suitable for exploring the role of Golgi-associated proteins in cancer progression.
GORASP1 (also known as GRASP65) encodes a peripheral Golgi membrane protein essential for Golgi cisternal stacking and ribbon formation. Mechanistically, GORASP1 forms a complex with GM130 (GOLGA2) to maintain Golgi architecture. During mitosis, this complex is regulated by phosphorylation through Cdk1 and Plk1, leading to Golgi disassembly, reversed upon dephosphorylation. Beyond its structural role, GORASP1 functions downstream of ERK, JNK, and mTOR, and governs integrin trafficking and secretion of extracellular matrix proteins, thereby influencing cell adhesion and migration. Additional interacting partners include USO1/p115, Dlg1, ??-catenin, and acetylated tubulin, underscoring its integration into pathways controlling cell morphology and motility.
In the HT29 colorectal adenocarcinoma background, disruption of GORASP1 provides a powerful system to dissect how Golgi integrity impacts malignant phenotypes. Given HT29 cells?? relevance to colorectal cancer, this knockout model is instrumental for examining GORASP1-dependent alterations in Golgi morphology, protein secretion, and cell adhesion/migration programs that drive metastasis. The polyclonal nature of the population allows researchers to assess the overall functional consequences of GORASP1 loss, including effects on mucin secretion and epithelial barrier function, and to screen compounds that may target Golgi-dependent pathways in cancer therapy.
This knockout cell product is suited for a wide range of experimental applications. Researchers can employ immunofluorescence staining to visualize Golgi fragmentation, western blotting and RT-qPCR to confirm GORASP1 ablation and downstream effects, and functional assays such as wound healing migration assays, transwell invasion assays, and cell adhesion assays to quantify motility changes. Additional utility includes ELISA-based secretion profiling, cell viability assays following drug treatment, and drug sensitivity screening for Golgi-targeted therapeutics. For further information or technical assistance, please contact Ascent Research.
