The KIF1C Knockout HT29 Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human HT29 colorectal adenocarcinoma line, in which the KIF1C gene has been disrupted. This genetically mixed population provides a loss-of-function model for studying the roles of the plus-end-directed microtubule motor protein KIF1C in cellular processes such as vesicle transport, adhesion dynamics, and migration, without the clonal variation associated with single-cell-derived lines. The polyclonal nature ensures representation of multiple independent editing events, offering a robust platform for functional genomics and cancer biology research.
HT29 is a widely used adherent epithelial cell line originally isolated from a 44-year-old female with colorectal adenocarcinoma. These cells exhibit characteristic epithelial morphology and retain many features of intestinal epithelium, including the capacity to form tight junctions and polarized monolayers, making them valuable for studying intestinal epithelial barrier function and colorectal cancer progression. The HT29 line is a well-established model for cancer biology, drug screening, and signaling studies, particularly in the context of adhesion, migration, and invasion??processes intimately linked to KIF1C function.
KIF1C is a kinesin-3 family motor that moves processively toward microtubule plus ends, transporting integrin-containing vesicles and other cargoes to the cell periphery. Its activity is regulated by integrin engagement, Src family kinases, 14-3-3 binding, and RhoA activation. KIF1C directly interacts with 14-3-3 proteins (YWHAB, YWHAZ), alpha-tubulin, integrin beta1, and kinesin light chain, and functions within a signaling network involving integrin alphaV/beta3, Src, FAK, paxillin, RhoA, and ROCK. Downstream, KIF1C promotes focal adhesion turnover, integrin recycling, MMP14 secretion, and cell migration speed. The mechanistic model indicates that KIF1C-mediated delivery of integrins to adhesion sites is critical for disassembly and reassembly of focal adhesions, enabling efficient cell motility.
In the HT29 colorectal cancer model, KIF1C knockout disrupts integrin trafficking, leading to reduced focal adhesion dynamics and impaired migratory and invasive capacity. This phenotype is particularly relevant for colorectal cancer metastasis, where tumor cell dissemination relies on coordinated adhesion turnover and matrix degradation. Additionally, KIF1C mutations are linked to hereditary spastic paraplegia type 58, underscoring its importance in neuronal transport; however, its role in epithelial cancer cell motility highlights a context-dependent vulnerability that can be explored using this knockout model. The system allows dissection of KIF1C??s contributions to colorectal cancer pathophysiology and its potential as a therapeutic target.
Researchers can employ these polyclonal KIF1C knockout HT29 cells in a range of assays to investigate cell migration, invasion, and cytoskeletal organization. Representative applications include Transwell migration and Matrigel invasion assays to quantify motility and invasiveness, immunofluorescence staining for focal adhesion markers such as vinculin and paxillin to assess adhesion dynamics, western blot analysis of KIF1C and downstream effectors (e.g., FAK, Src, MMP14), RT-qPCR profiling of epithelial-mesenchymal transition markers, and adhesion assays on extracellular matrix substrates. These tools support studies in cancer cell biology, cytoskeletal dynamics, motor protein function, and drug target validation. For additional technical details, please contact Ascent Research.
