The GORASP2 Knockout HT29 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population for loss-of-function studies of GORASP2. This pooled population, derived from the HT29 human colorectal adenocarcinoma line, carries targeted disruptions of the GORASP2 gene, thereby eliminating functional GORASP2 protein expression. The polyclonal nature enables robust experimental replication across a heterogeneous cell pool without clonal selection artifacts.
HT29 is a widely used epithelial cell line established from a primary colorectal adenocarcinoma of a 44-year-old female. These cells are instrumental in investigating intestinal epithelial biology, including differentiation, barrier function, and drug transport, and serve as a standard in vitro model for colorectal cancer research. Their retained epithelial characteristics and ease of culture make them a reliable platform for dissecting molecular mechanisms underlying colon carcinoma progression and metastasis.
GORASP2 encodes a Golgi reassembly stacking protein that is essential for mitotic Golgi fragment reunification and post-mitotic cisternal restacking. Beyond its structural role, GORASP2 functions in autophagy by tethering LC3-positive autophagosomes via direct interaction with the LC3 family (MAP1LC3A/B), facilitating lysosomal fusion. In cell migration, GORASP2 regulates integrin ??1 (ITGB1) trafficking to the cell surface through formation of a complex with paxillin (PXN), thereby controlling adhesion turnover. GORASP2 activity is phosphorylated by CDK1 and PLK1 during mitosis, and its autophagy function is regulated downstream of ULK1 and mTORC1, through interactions with GM130 (GOLGA2) and GORASP1.
In the HT29 colorectal cancer context, GORASP2 knockout provides a powerful model to dissect its contributions to Golgi integrity, autophagic flux, and migratory capacity that are often dysregulated in tumor progression. The disruption of GORASP2 allows researchers to examine how Golgi structural defects impact secretory pathway function, and to evaluate the consequences on integrin-mediated cell adhesion and directional movement, processes critical for metastatic dissemination of colorectal carcinoma.
Researchers can employ these polyclonal knockout cells in a broad array of assays, including immunofluorescence microscopy for Golgi markers (GM130, Giantin) and autophagosome puncta (LC3), western blotting for LC3-II and p62 turnover, wound healing and transwell migration assays, co-immunoprecipitation to map protein interaction networks, and electron microscopy for ultrastructural analysis of Golgi morphology. The cells are also suitable for high-content screening platforms aimed at identifying modulators of Golgi reassembly or autophagy. For further technical details and ordering information, please contact Ascent Research.
