KIAA0319L Knockout A-549 Polyclonal Cells offer a polyclonal population of A-549 human lung adenocarcinoma cells with CRISPR/Cas9-mediated disruption of the KIAA0319L gene. This gene encodes the adeno-associated virus receptor (AAVR), which is essential for viral entry. The knockout is generated using a non-homogeneous editing approach, resulting in a mixed pool of cells with various loss-of-function alleles, providing a model to study KIAA0319L function without clonal selection. The product is supplied as a polyclonal population, enabling robust representation of diverse genetic backgrounds.
A-549 cells are a human lung adenocarcinoma cell line derived from a 58-year-old Caucasian male with pulmonary carcinoma. They exhibit adherent epithelial morphology and serve as a canonical model for type II alveolar epithelial cells. Widely used in cancer biology, these cells are instrumental for investigating oncogenic signaling, drug metabolism, and host-pathogen interactions, particularly respiratory viruses. Their well-characterized transcriptome and ease of manipulation make them suitable for CRISPR-based genome editing and functional studies.
KIAA0319L (AAVR) acts as a primary receptor for multiple AAV serotypes, directly binding viral capsids to trigger clathrin-mediated endocytosis. This process involves the AP2 adaptor complex and dynamin for vesicle formation and scission. Internalized virions traffic through early endosomes marked by Rab5 and late endosomes regulated by Rab7, ultimately leading to endosomal escape and nuclear import of the AAV genome for transduction. KIAA0319L is thus essential for efficient viral entry and intracellular trafficking. Its expression in A-549 cells appears constitutive, controlled by general transcription factors, and no direct transcriptional targets of KIAA0319L are known; instead, its function is mediated through physical interactions with endocytic machinery and AAV capsids.
In A-549 lung carcinoma cells, KIAA0319L knockout provides a physiologically relevant platform to dissect AAV entry mechanisms in a pulmonary epithelial environment. Given the lung’s importance as a target for AAV-based gene therapies, this model enables the study of AAV serotype tropism and transduction efficiency in cancer-derived lung cells. The absence of KIAA0319L creates a powerful AAV-resistant control, which is essential for validating vector specificity, off-target effects, and receptor dependency in gene delivery experiments. Furthermore, coupling this knockout with the A-549 background allows exploration of how malignant transformation might influence AAV receptor expression and viral internalization pathways.
Researchers can employ these polyclonal knockout cells in a variety of quantitative assays, including Western blotting and RT-qPCR to confirm KIAA0319L ablation, and flow cytometry to measure surface receptor loss and impaired AAV binding. Transduction reporter assays using GFP- or luciferase-encoding AAV vectors directly quantify the functional consequence of KIAA0319L disruption. Additionally, clathrin-mediated endocytosis can be interrogated using pharmacological inhibitors in conjunction with viral uptake assays. These cells are instrumental for AAV transduction mechanism studies, gene therapy vector development, and pulmonary gene delivery modeling. For further details or technical support, please contact Ascent Research.
