The KIF1C Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed for the disruption of the KIF1C gene in the HAP1 human cell line. This product provides a heterogeneous pool of cells carrying targeted gene disruption, enabling robust functional studies of KIF1C-dependent processes without the need for clonal isolation. The polyclonal format maintains genetic diversity while ensuring loss-of-function across the majority of cells.
The HAP1 cell line is an adherent, near-haploid human fibroblast-like line derived from KBM-7 cells, originally isolated from a male patient with chronic myeloid leukemia in blast crisis. Its haploid karyotype simplifies the generation of knockout models, as a single targeting event can eliminate gene function without interference from a second allele. This feature makes HAP1 an ideal platform for high-throughput genetic screening and detailed phenotypic characterization, particularly in studies of cell adhesion, migration, and cytoskeletal organization.
KIF1C encodes a microtubule-based plus-end-directed motor protein of the kinesin-3 family that is essential for the intracellular transport of integrin-containing vesicles and other cargo. It is regulated by upstream factors such as the Rab6 GTPase?CBicaudal D cargo adaptor complex, phosphorylation by protein kinase A (PKA), and binding to 14-3-3 proteins and the tyrosine phosphatase PTPN21. KIF1C directly mediates the delivery of integrins (??5??1, ??v??3) to focal adhesions, affecting the localization and turnover of paxillin and vinculin, and influencing actin polymerization and Rho GTPase activity. Interacting partners include Hook1 and the dynactin complex via PTPN21, which couples KIF1C to the microtubule network. Through these interactions, KIF1C orchestrates focal adhesion dynamics and cell migration.
In HAP1 cells, which intrinsically display strong adhesive and migratory properties, KIF1C disruption generates a powerful model to dissect the molecular mechanisms of integrin trafficking and adhesion turnover. The polyclonal knockout population captures a range of mutation events, potentially revealing hypomorphic or compensatory effects, while the haploid background ensures clear loss-of-function phenotypes. This model is particularly suited to examining the interplay between microtubule-based transport and actin cytoskeleton remodeling during cell spreading and directional movement.
These knockout cells can be employed in a variety of assays, including scratch wound healing and Transwell migration/invasion assays to quantify cell motility defects, live-cell imaging of fluorescently tagged vesicle cargo to monitor transport dynamics, and immunofluorescence staining for focal adhesion components (vinculin, paxillin) and phalloidin for actin organization. Applications span neurodegeneration research (modeling SPG58), cancer biology (assessing metastatic potential), and drug discovery (screening for modulators of kinesin motor proteins). For detailed protocols, lot-specific characterization, and technical support, please contact Ascent Research.
