The KIFC3 Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the A-549 human lung adenocarcinoma cell line, with targeted disruption of the KIFC3 gene. This product provides a loss-of-function model for investigating the minus-end-directed kinesin motor protein KIFC3, which participates in microtubule-dependent vesicle transport and mitotic spindle dynamics. The polyclonal nature avoids clonal bias and represents a heterogeneous knockout pool suitable for population-level functional assays.
The parental A-549 cell line is a widely used adherent epithelial model originating from human lung adenocarcinoma tissue, displaying a type II alveolar epithelial phenotype. These cells are extensively characterized in cancer biology, serving as a standard system for studying lung adenocarcinoma signaling, proliferation, and drug responses. Their retention of alveolar epithelial features makes them particularly relevant for examining intracellular trafficking and cytoskeletal organization in a disease context.
KIFC3 belongs to the kinesin-14 family and functions as a minus-end-directed microtubule motor. It interacts directly with the dynein-dynactin complex and is critical for the transport of Rab11-FIP3-positive recycling endosomes. KIFC3 activity is regulated during cell division by upstream kinases CDK1-cyclin B and PLK1, and it contributes to spindle pole focusing by mediating microtubule sliding. Downstream effects involve Rab11 and FIP3 vesicle cargo delivery, with crosstalk to importin beta. Representative pathway components include KIFC3, dynein/dynactin, Rab11, FIP3, CDK1, and PLK1. Loss of KIFC3 disrupts endosomal recycling and may impair mitotic spindle organization due to altered microtubule dynamics.
In the A-549 adenocarcinoma context, KIFC3 knockout provides a physiologically relevant platform for studying how kinesin motor defects impact cancer cell division and vesicle trafficking. KIFC3 dysfunction is associated with mitotic defects and may sensitize cells to spindle-targeting agents, making this model valuable for anticancer drug screening. The polyclonal knockout pool reflects heterogeneous responses that better mimic tumor heterogeneity compared to single-cell clones, enabling robust analysis of kinesin-driven processes in lung cancer biology.
Applications include live-cell imaging of Rab11-FIP3 endosomal trafficking, immunofluorescence microscopy of ??-tubulin to assess spindle morphology, and western blotting to confirm KIFC3 ablation. Functional assays such as flow cytometry for cell cycle distribution, MTS/MTT proliferation assays, and Transwell migration assays are also commonly performed. This knockout population supports mechanistic studies of kinesin-14 motors, microtubule-dependent transport, and the identification of mitotic inhibitors in lung adenocarcinoma research. For additional details, contact Ascent Research.
