This product is a CRISPR/Cas9-edited polyclonal knockout cell population derived from A-549 lung adenocarcinoma cells, with targeted disruption of the BRD8 gene. The polyclonal nature provides a heterogeneous pool of edited alleles, suitable for population-level assays and pooled screening without clonal isolation. The knockout model results in functional loss of BRD8 protein, enabling investigation of BRD8-dependent processes.
A-549 cells, established from a lung adenocarcinoma patient, are hypotriploid epithelial cells retaining features of type II pneumocytes. They harbor an activating KRAS G12S mutation and express wild-type EGFR, making them a key model for non-small cell lung cancer research. Their extensive characterization in drug response and signaling studies provides a reliable background for gene perturbation experiments.
BRD8 is a scaffolding subunit of the NuA4/TIP60 histone acetyltransferase complex, where its bromodomain recognizes acetylated histone H4 to facilitate chromatin remodeling and transcription. Upstream regulators include E2F1, androgen receptor, and DNA damage signals. BRD8 interacts with TIP60/KAT5, EP400, and TRRAP, and downstream it promotes acetylation of p53 and histone H4, thereby activating transcription of E2F target genes such as CCND1 and CCNE1, as well as DNA repair genes. Through these interactions, BRD8 integrates signals to coordinate cell cycle progression, apoptosis, and genome stability. In addition, BRD8 has been implicated in the DNA damage response through its interaction with DNA-PKcs and its role in histone acetylation near double-strand breaks.
In A-549 cells, BRD8 knockout impairs NuA4-mediated chromatin remodeling, leading to reduced histone H4 acetylation and diminished p53-dependent transcriptional responses. This results in altered expression of E2F-driven cell cycle genes and compromised DNA damage repair, as indicated by increased ??-H2AX foci. Consequently, the model may exhibit altered proliferation, survival, and sensitivity to genotoxic agents, providing a system to study epigenetic dysregulation in lung adenocarcinoma progression and therapeutic resistance.
Applications include epigenetic mechanism studies in lung cancer, DNA damage repair pathway analysis, and drug sensitivity screening with cisplatin or etoposide. Common assays are Western blotting for acetylated H4 and p53, RT-qPCR for CCND1, immunofluorescence for ??-H2AX foci, flow cytometry for cell cycle, and clonogenic survival assays. Co-immunoprecipitation can evaluate NuA4 complex assembly, and chromatin immunoprecipitation can assess BRD8 chromatin binding. Transcriptomic analyses via RNA-seq can further define BRD8-dependent gene networks. This polyclonal knockout cell population is a valuable tool for functional genomics and preclinical cancer research. For further information, please contact Ascent Research.
