The KLC4 Knockout HT29 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal population derived from the human colorectal adenocarcinoma cell line HT29, with targeted disruption of the KLC4 gene. This product provides a loss-of-function model to investigate the cellular roles of kinesin light chain 4 (KLC4), a critical component of the kinesin-1 motor complex. These cells are designed for researchers aiming to dissect microtubule-dependent intracellular transport mechanisms in a cancer-relevant epithelial context.
The parental HT29 cell line was established from a primary colorectal adenocarcinoma of a 44-year-old Caucasian female. HT29 cells are widely employed as a model system for intestinal epithelial biology and colorectal cancer research, exhibiting adherent epithelial morphology and retaining key features of tumor cell behavior. This cell line is valuable for investigating pathways involved in carcinogenesis, metastasis, drug resistance, and cellular differentiation. Its well-characterized signaling networks and ease of manipulation make it an ideal host for CRISPR/Cas9-mediated gene disruption studies.
The KLC4 gene encodes a kinesin light chain that forms obligate heterotetramers with kinesin heavy chains, predominantly of the KIF5 subfamily (KIF5A, KIF5B, KIF5C), to assemble the kinesin-1 motor complex. KLC4 directly mediates cargo binding through interactions with adaptor proteins such as JNK-interacting protein 1 (JIP1) and participates in the transport of diverse vesicular cargoes, including endosomes and lysosomes, along microtubules. Upstream, KLC4 function is regulated by stress-activated kinases, notably JNK (MAPK8/9), which phosphorylate cargo adaptors to modulate motor?Ccargo coupling. Downstream, kinesin-1 motors transport components critical for focal adhesion dynamics and JNK signaling, implicating KLC4 in the spatiotemporal control of cell migration and stress responses.
In the context of colorectal adenocarcinoma, disruption of KLC4 is expected to perturb the microtubule-dependent trafficking essential for maintaining epithelial polarity, directional migration, and signaling compartmentalization. HT29 cells depend on cytoskeletal dynamics for invasive behavior and survival, making this knockout model particularly relevant for examining how transport defects influence cancer progression. By impairing the delivery of key molecules to specific subcellular domains, KLC4 knockout may alter JNK pathway activity, focal adhesion turnover, and vesicular recycling, thereby affecting cell?Cmatrix interactions and metastatic potential.
These polyclonal KLC4 knockout HT29 cells enable Western blotting to confirm target protein loss, immunofluorescence and live-cell imaging to track organelle distribution and vesicle motility, and functional assays??including wound healing, transwell migration, viability, and apoptosis??to evaluate cell behavior and drug sensitivity. Subcellular fractionation assists in detecting cargo mislocalization and altered signaling complex assembly. This model thus offers a versatile platform for dissecting transport-dependent regulation of colorectal cancer phenotypes. For technical inquiries or ordering, please contact Ascent Research.
