BRPF3 Knockout A-549 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human A-549 lung carcinoma cell line, engineered to disrupt the BRPF3 gene and abolish protein expression. This loss-of-function model provides a heterogeneous pool of cells with targeted gene disruption, enabling robust functional genomics studies without the selective pressures of clonal isolation. The knockout is achieved through CRISPR/Cas9-mediated non-homologous end joining, generating a population-level ablation of BRPF3 function that facilitates investigation of its role in chromatin regulation and cell cycle control within a non-small cell lung cancer (NSCLC) context.
The parental A-549 cell line originates from lung adenocarcinoma tissue of a 58-year-old Caucasian male and exhibits adherent, epithelial-like morphology. A-549 cells are widely recognized as a model for NSCLC, extensively used to study cancer biology, oncogenic signaling, drug responses, and metastatic mechanisms. Their genetic background, including mutant KRAS, drives constitutive proliferation, making them particularly suitable for dissecting the contributions of epigenetic regulators such as BRPF3 to tumorigenesis. This knockout thus provides a valuable tool for examining chromatin-mediated control in lung adenocarcinoma.
BRPF3 functions as a scaffold protein that assembles the HBO1 (KAT7) histone acetyltransferase complex, interacting with HBO1, ING4/5, and MEAF6 to direct acetylation of histone H4 at lysines 5, 8, and 12 (H4K5ac, H4K8ac, H4K12ac). This acetylation relaxes chromatin, promoting transcriptional activation of cell cycle genes including CCND1 and MYC, and licensing DNA replication origins via regulation of the MCM complex. Consequently, BRPF3 promotes cell cycle progression and proliferation. Knockout of BRPF3 disrupts complex formation, leading to reduced H4 acetylation, impaired expression of downstream proliferation drivers, and attenuation of cell cycle entry.
In the A-549 NSCLC model, loss of BRPF3 is poised to compromise the histone acetylation landscape that sustains oncogenic transcription and replication. Given the HBO1 complex’s role in maintaining epigenetic plasticity, BRPF3 knockout cells serve as a platform to investigate acetylation-dependent chromatin remodeling contributions to lung adenocarcinoma pathogenesis. This model enables exploration of synthetic lethal interactions, assessment of epigenetic drug sensitivities, and identification of molecular dependencies, potentially revealing vulnerabilities associated with deficient H4 acetylation in NSCLC.
Researchers can employ BRPF3 Knockout A-549 Polyclonal Cells in a range of assays to probe epigenetic regulation and cell cycle control. Representative techniques include western blotting for H4K5ac, H4K8ac, and H4K12ac; RT-qPCR for CCND1 and MYC transcript levels; ChIP-qPCR to map histone acetylation at replication origins; co-immunoprecipitation to assess HBO1 complex assembly; and functional assays such as MTT proliferation, BrdU incorporation, flow cytometric cell cycle analysis, and colony formation. These cells are suitable for genetic complementation, high-content screening of epigenetic inhibitors, and mechanistic dissection of the HBO1 acetyltransferase axis. For further details and technical inquiries, please contact Ascent Research.
