AKNA Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal human lung epithelial cell population with targeted disruption of the AKNA gene. The product comprises a heterogeneous pool of A-549 cells harboring gene edits introduced by non-homologous end joining-mediated repair, resulting in loss of functional AKNA protein expression. This knockout model enables investigation of AKNA-dependent transcriptional programs without clonal selection, preserving population-level responses relevant to epithelial biology.
A-549 cells are an adherent human alveolar basal epithelial cell line originally derived from lung adenocarcinoma tissue of a 58-year-old Caucasian male. These cells exhibit key characteristics of type II pneumocytes, including lamellar bodies and surfactant production, and are widely used to model pulmonary epithelial barrier function, innate immune responses, and lung adenocarcinoma biology. Their epithelial origin and robust growth make them suitable for genetic manipulation and functional assays.
AKNA encodes an AT-hook transcription factor that orchestrates immune and inflammatory gene expression, epithelial differentiation, and mucosal barrier integrity. In response to upstream signals such as TNF-alpha, IFN-gamma, and LPS/TLR4 engagement, AKNA is activated and forms complexes with cofactors including NF-kappa-B p65, IRF1, and p300/CBP to regulate downstream targets. Through these interactions, AKNA promotes transcription of cytokines (IL6, IL8, TNFA), antigen-presentation molecules (MHC-I/II), and tight junction components (CLDN1, OCLN, TJP1), while integrating signals from NF-??B, STAT1, and TLR pathways. Disruption of AKNA abrogates these regulatory networks.
Given the central role of A-549 cells in pulmonary research, AKNA knockout in this background provides a clinically relevant system to dissect mechanisms of airway inflammation and barrier dysfunction. Loss of AKNA impairs NF-??B and interferon-mediated transcriptional responses, attenuating cytokine secretion and compromising epithelial tight junction integrity. This mimics aspects of inflammatory lung diseases such as asthma and lung adenocarcinoma progression, where AKNA dysregulation has been implicated. The polyclonal nature of the knockout pool mirrors the genetic heterogeneity often observed in tumor environments, offering a realistic model for drug intervention studies.
Researchers can employ this knockout model to study immune and inflammatory gene regulation, epithelial barrier function, infection biology (viral/bacterial), and lung cancer pathogenesis. Typical assays include Western blotting for AKNA and IL6, RT-qPCR for cytokine panels, immunofluorescence for NF-??B p65 translocation, TEER measurements for barrier integrity, ELISA for IL6/8, and ChIP-qPCR for AKNA binding. Additionally, flow cytometry for MHC-I/II and co-immunoprecipitation of the AKNA interactome extend the model’s utility. For further technical specifications, please contact Ascent Research.
