KIFC3 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HAP1 line, carrying targeted disruption of the KIFC3 gene. Generated without single-cell cloning, this pooled knockout product captures diverse loss-of-function mutations, minimizing clonal artifacts and enabling robust population-level analyses. The polyclonal format is particularly suited for functional genomics screens and studies of heterogeneous knockout phenotypes in a near-haploid background.
The HAP1 host cell line is an adherent, near-haploid human cell line with fibroblast-like morphology, originally derived from a male chronic myelogenous leukemia patient. Its haploid karyotype??with a single copy of most chromosomes??facilitates efficient gene disruption and ensures that single-allele knockout yields potent loss-of-function phenotypes. HAP1 cells maintain intact PI3K/AKT and mTOR signaling pathways, making them a valuable model for cancer cell biology, haploid genetic screens, and drug mechanism-of-action studies.
KIFC3 encodes a minus-end-directed kinesin motor protein that powers retrograde transport along microtubules, primarily responsible for Golgi apparatus organization and vesicle trafficking. Mechanistically, KIFC3 forms complexes with Golgin-160 and dynein components to tether vesicles and maintain Golgi stack integrity. Its expression is post-transcriptionally regulated by miR-200c. Downstream, KIFC3 activity promotes AKT phosphorylation, activating the PI3K/AKT/mTOR signaling axis that drives cell cycle progression and suppresses apoptosis. In cancer cells, KIFC3 upregulation reinforces these proliferative signals and contributes to spindle assembly checkpoint accuracy. Consequently, KIFC3 loss disrupts Golgi morphology, attenuates AKT signaling, and impairs mitotic fidelity.
In the HAP1 context, the near-haploid genome ensures that KIFC3 knockout elicits clear and unconfounded phenotypes, making it an excellent system for investigating Golgi-dependent trafficking and mitotic functions. The chronic myelogenous leukemia origin directly links this model to leukemogenesis studies, while emerging evidence implicates KIFC3 in hepatocellular carcinoma, breast cancer, and gastric cancer, broadening its relevance to solid tumor biology. The polyclonal knockout population also provides a heterogeneous background useful for assessing drug tolerance and adaptive responses.
Researchers can employ these cells in a variety of assays: Western blotting and RT-qPCR verify KIFC3 knockout efficiency; immunofluorescence microscopy visualizes Golgi fragmentation and spindle defects; flow cytometry enables cell cycle and apoptosis profiling; and co-immunoprecipitation with Golgin-160 or dynein subunits confirms disrupted interactions. Migration and invasion assays gauges metastatic potential, while drug sensitivity screens with PI3K/AKT/mTOR inhibitors probe compensatory signaling. Genome-wide haploid screens can identify synthetic lethal interactions. For detailed specifications or custom inquiries, please contact Ascent Research.
