The KIF16B Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from the HAP1 cell line, engineered for disruption of the KIF16B gene. This gene-edited product provides a loss-of-function model for studying the kinesin motor protein KIF16B in a near-haploid human background. The polyclonal knockout format ensures representation of a variety of editing events across the cell population, enabling robust functional genomic studies without the need for clonal isolation. The cells are intended for receptor trafficking, signal transduction, and oncology research applications.
HAP1 is a near-haploid human cell line originally derived from a male patient with chronic myeloid leukemia (CML). Its haploid karyotype makes it an ideal host for CRISPR-based functional genomics, as single-copy gene disruption yields clear phenotypic readouts. Widely employed in genome-wide knockout screens, HAP1 cells retain key signaling pathways relevant to cancer biology, including EGFR-MAPK and PI3K-AKT cascades. This background provides a simplified yet physiologically relevant platform for dissecting gene function, particularly in kinase signaling and vesicle trafficking networks.
KIF16B encodes a kinesin-3 family motor that directs plus-end transport of early endosomes along microtubules. Its PX domain binds phosphatidylinositol 3-phosphate (PtdIns(3)P) on endosomes, linking motility to phosphoinositide signaling downstream of Vps34/PI3K-III and Rab5. KIF16B critically regulates recycling of internalized receptors, notably epidermal growth factor receptor (EGFR), to the plasma membrane, sustaining MAPK/ERK and AKT pathway activity. It interacts with early endosome components including Rab5, Vps34, p150, and EEA1. By modulating receptor trafficking, KIF16B influences cell migration, proliferation, and invasion.
Disruption of KIF16B in the HAP1 background creates a powerful model to interrogate endosomal trafficking and its impact on oncogenic signaling. The near-haploid genotype ensures that phenotypic consequences are directly attributable to KIF16B loss, without confounding effects from wild-type alleles. This knockout model is particularly relevant for glioblastoma and breast cancer research, where KIF16B has been implicated in tumor cell migration and invasion. Researchers can use this system to explore how receptor sorting defects alter downstream signaling networks and cellular behaviors, including drug sensitivity to EGFR-targeted therapies.
The KIF16B Knockout HAP1 Polyclonal Cells are suitable for an array of experimental applications. Receptor recycling dynamics can be assessed by flow cytometry to quantify surface EGFR levels, while immunofluorescence microscopy can visualize endosome distribution. Downstream signaling activity is readily measured by western blotting or phospho-specific analysis for ERK, AKT, and other pathway components. Functional assays such as cell proliferation, migration, and invasion assays can reveal the consequences of disrupted KIF16B-dependent trafficking. Additionally, these cells can be employed in drug sensitivity screens with EGFR inhibitors to investigate resistance mechanisms. For further details and support, please contact Ascent Research.
