The KIF21A Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from the HT29 human colorectal adenocarcinoma cell line. This product provides a mixed population of knockout cells with targeted disruption of the Kinesin Family Member 21A (KIF21A) gene, generated using CRISPR/Cas9-mediated gene editing. The polyclonal format offers a heterogeneous pool of edited alleles, enabling the study of gene function without the need for single-cell cloning. This model serves as a versatile tool for dissecting microtubule-dependent processes in epithelial cancer cells.
The parental HT29 cell line originates from a 44-year-old female patient with colorectal adenocarcinoma and exhibits an epithelial, moderately differentiated phenotype. HT29 cells are widely employed in cancer research, particularly for studies on colon adenocarcinoma biology, cellular adhesion, and epithelial-to-mesenchymal transition. Their robust growth and well-characterized signaling networks make them a reliable host for gene knockout studies, facilitating the investigation of cytoskeletal dynamics and intracellular trafficking in a malignant epithelial context.
KIF21A encodes a plus-end-directed microtubule motor protein transporting cargoes along axons in neurons and contributing to microtubule organization in non-neuronal cells. Its motor activity is regulated by upstream factors including neuronal growth factor NGF, intracellular calcium signaling, and microtubule-associated regulatory kinases. KIF21A interacts with kinesin light chains (KLCs), the KIFBP adaptor protein, and the dynactin complex to form transport complexes. Upon activation, it binds microtubules and hydrolyzes ATP to move downstream cargoes such as synaptic vesicle precursors and cell adhesion molecules along the cytoskeleton. This cascade integrates microtubules, ATP, KLCs, KIFBP, and the neuronal cytoskeleton to coordinate transport and architecture.
In the HT29 colorectal adenocarcinoma background, knockout of KIF21A disrupts intracellular trafficking and cytoskeletal integrity, potentially affecting cell morphology, proliferation, and migration. This model enables dissection of kinesin-driven microtubule dynamics in epithelial cancer cell biology. Although KIF21A mutations are primarily linked to congenital fibrosis of the extraocular muscles type 1 (CFEOM1) and neurodevelopmental disorders, its knockout in HT29 cells allows exploration of fundamental microtubule-dependent mechanisms relevant to neuronal and non-neuronal pathologies. The polyclonal population models tumor cell heterogeneity, offering insights into how loss of KIF21A influences collective cellular behavior.
Typical applications include Western blotting, RT-qPCR, and immunofluorescence to validate KIF21A disruption and assess microtubule organization. Functional assays such as cell migration and proliferation assays are employed to evaluate phenotypic consequences. This product is suited for studying kinesin function in colorectal cancer, investigating microtubule-related processes in epithelial cells, and screening for CFEOM1-associated mechanisms. Researchers seeking to elucidate intracellular transport pathways or test therapeutic interventions targeting the microtubule cytoskeleton will find this model a valuable resource. For additional information or technical support, please contact Ascent Research.
