The KIF13A Knockout HT29 Polyclonal Cells product consists of a CRISPR/Cas9-edited HT29 cell pool with targeted disruption of the motor protein KIF13A, yielding a heterogeneous knockout population. This format provides a robust tool for loss-of-function studies without the need for clonal isolation.
HT29 cells are a well-established human colorectal adenocarcinoma line derived from a primary tumor. These epithelial cells can differentiate into enterocyte-like cells under appropriate conditions, making them a relevant model for intestinal epithelial biology and colorectal cancer research. The expression of integrins, cadherins, and endosomal trafficking components in HT29 cells makes them a suitable host for examining KIF13A functions in adhesion and vesicle transport.
KIF13A is a plus-end-directed kinesin-3 motor that drives transport of endosomal cargo to the cell periphery. It directly interacts with the adaptor RILP, the BLOC-1 complex, and centaurin-??1 (ADAP1), and its activity is regulated by phosphatidylinositol 3-phosphate (PI3P) and the GTPase Rab7. Through this network, KIF13A governs recycling of ??1 integrin-containing vesicles to the plasma membrane and positions lysosomes via Rab7?CRILP effectors. These functions control focal adhesion dynamics, cell migration, and endolysosomal degradative capacity.
In HT29 colorectal adenocarcinoma cells, KIF13A-driven ??1 integrin recycling is critical for maintaining cell-substrate adhesion and supporting collective and single-cell migration. Knockout of KIF13A disrupts these functions, enabling dissection of endosomal trafficking mechanisms that underlie cancer cell invasion and metastasis. Lysosome mispositioning also alters mTORC1 nutrient sensing and chemosensitivity, while KIF13A??s involvement in hepatitis C virus replication broadens applicability to virology research.
Researchers can employ these polyclonal knockout cells in Western blotting, RT-qPCR, and co-immunoprecipitation to confirm gene disruption and assess binding partners. Immunofluorescence and LysoTracker staining visualize lysosome distribution, while Transwell migration and wound healing assays measure cell motility. Flow cytometry quantifies surface ??1 integrin, and the model supports drug testing where endolysosomal pathways affect therapeutic response. For further information, please contact Ascent Research.
