The KIF20B Knockout HEK293T Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HEK293T host cell line, designed to disrupt expression of the KIF20B gene. This cell pool provides a heterogeneous loss-of-function model for studying the roles of the kinesin motor protein KIF20B in cell division and related processes. The polyclonal format captures a broad spectrum of editing outcomes without single-cell cloning, making it suitable for pooled functional screens and robust phenotype analysis.
The HEK293T cell line is a widely used human embryonic kidney epithelial cell model engineered with the SV40 large T antigen, enabling episomal replication of transfected plasmids containing the SV40 origin of replication. These cells exhibit high transfection efficiency and robust proliferation, making them a standard platform for protein expression, viral packaging, and gene editing studies. Their epithelial origin and rapid growth provide a relevant context for examining genes involved in cell cycle regulation and mitotic progression.
KIF20B encodes a plus-end-directed kinesin motor protein essential for cytokinesis. During late mitosis, KIF20B accumulates at the central spindle and transports Aurora B kinase to the midzone, facilitating the activation of the RhoA signaling cascade. This includes the recruitment of MgcRacGAP, Ect2, and RhoA itself, leading to assembly of the actomyosin contractile ring via effectors such as Citron, Anillin, and Myosin II. KIF20B is regulated by CDK1, Aurora B, PLK1, and the transcription factor FOXM1, and it interacts with mitotic regulators including INCENP, Survivin, and CEP55. Disruption of KIF20B impairs central spindle assembly and cleavage furrow ingression, resulting in failed cytokinesis and genome instability.
In the HEK293T background, KIF20B knockout disrupts normal cell division, potentially leading to multinucleation, aneuploidy, and altered proliferation dynamics. This model is particularly valuable for dissecting the molecular requirements of cytokinesis and exploring the consequences of mitotic failure in a highly tractable cell line. Given the frequent dysregulation of mitotic kinesins and chromosomal instability in cancer, these cells offer a relevant system for investigating KIF20B??s role in tumor cell biology and characterizing dependencies on the RhoA?Ccytokinesis pathway.
Key applications include detailed phenotypic analysis using immunofluorescence to monitor spindle morphology and Aurora B localization, Western blotting for KIF20B and phospho-histone H3 levels, and flow cytometry to assess DNA content and cell cycle distribution. Live-cell imaging enables real-time observation of mitotic progression, while co-immunoprecipitation can identify interaction partners such as Aurora B or PLK1. Additionally, these cells support proliferation and viability assays for drug screening and migration/invasion studies relevant to cancer metastasis. For further technical information or bulk orders, please contact Ascent Research.
