The BRWD1 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited knockout cell population derived from HEK293T human embryonic kidney epithelial cells. This polyclonal pool provides a loss-of-function model for studying BRWD1-dependent processes in chromatin organization, transcriptional regulation, and DNA damage repair. The heterogeneous nature of the population reduces clonal bias and facilitates robust functional genomics studies.
HEK293T cells are an immortalized subclone of HEK293 cells stably expressing the SV40 large T antigen, which enhances plasmid replication and protein expression. Their high transfectability and well-defined biology make them a standard host for transient transfection, protein production, and signaling studies. As kidney epithelial cells, they retain relevant cellular machinery for investigating chromatin dynamics and cell cycle control.
BRWD1 is a chromatin reader with bromodomains that recognize acetylated histones, enabling it to scaffold chromatin remodeling complexes such as the NuA4 acetyltransferase complex (including EP400 and TRRAP) and the SWI/SNF ATPase complex (SMARCA4 and SMARCC1). It acts downstream of Notch signaling via NICD and is regulated by TP53 and E2F transcription factors, positioning BRWD1 at the intersection of G1/S cell cycle transition and DNA double-strand break repair. BRWD1 transcriptional targets include RAG1 and RAG2, and it modulates chromatin accessibility at gene promoters; disruption thus impairs the function of these remodeling complexes and downstream gene expression programs.
In HEK293T cells, BRWD1 knockout provides a suitable model to examine conserved chromatin regulatory mechanisms outside of lymphoid lineages. Although BRWD1 is critical for B cell development and V(D)J recombination, its core activities in histone acetylation and SWI/SNF-mediated remodeling are relevant to epithelial cells. This enables investigation of BRWD1’s role in DNA damage response and cell cycle progression in a genetically tractable background compatible with high-throughput assays.
Applications include functional analysis of chromatin organization, validation of Notch pathway interactions, and exploration of DNA repair pathways. Assays such as ChIP-qPCR for histone modifications, co-immunoprecipitation of BRWD1 complexes, RNA-seq for transcriptome profiling, comet assay for DNA damage, and cell cycle analysis by flow cytometry can be performed. The model also supports drug target validation for B-cell lymphoma and immunodeficiency research. For more details, contact Ascent Research.
