The BRPF3 Knockout HT29 Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 colorectal adenocarcinoma cell line, featuring targeted disruption of the BRPF3 gene. This loss-of-function model provides a heterogeneous genetic background, capturing a broad spectrum of knockout-associated effects without the bias of single-cell clonal selection. It serves as a versatile research tool for investigating BRPF3-dependent histone acetylation dynamics and chromatin remodeling in the context of colorectal cancer biology.
HT29 cells were originally isolated from a primary colon adenocarcinoma of a 44-year-old female and are extensively utilized as an epithelial model for colorectal cancer research. These adherent cells harbor well-characterized mutations in the tumor suppressor genes APC and TP53, as well as oncogenic alterations in KRAS and PIK3CA, thereby recapitulating key molecular features of colorectal tumorigenesis. Additionally, HT29 cells retain the capacity for enterocytic differentiation upon appropriate stimulation, allowing the study of differentiation-associated epigenetic changes when BRPF3 function is disrupted.
BRPF3 encodes a scaffold protein integral to the HBO1 (KAT7) histone acetyltransferase complex, where it physically links the catalytic subunit HBO1 to chromatin and directs the acetylation of histone H3 and H4 tails. This post-translational modification relaxes chromatin structure, enabling transcriptional activation of genes essential for DNA replication, such as the pre-replication complex components CDC6 and CDT1. Upstream regulators include E2F transcription factors and cell cycle kinases, which couple BRPF3 activity to proliferative signals. Within the complex, BRPF3 interacts with HBO1, ING5, EAF6, and the histone substrates themselves. Disruption of BRPF3 impairs assembly of the HBO1 complex, reduces global and locus-specific histone H3/H4 acetylation, and consequently perturbs gene expression programs and cell cycle progression.
In the HT29 colorectal adenocarcinoma model, BRPF3 knockout allows dissection of how compromised histone acetylation influences cancer cell behavior. The pre-existing mutations in this cell line already dysregulate key signaling pathways; loss of BRPF3 adds an epigenetic layer of perturbation, potentially exacerbating transcriptional defects or revealing synthetic vulnerabilities. This makes the model valuable for studying the interplay between histone modification pathways and oncogenic signaling, and for evaluating the functional significance of BRPF3 in sustaining the malignant phenotype.
This polyclonal knockout cell population supports a broad array of experimental applications. Western blotting with antibodies against total and acetylated histone H3/H4, combined with RT-qPCR for downstream targets such as CDC6 and CDT1, enables direct assessment of BRPF3-dependent acetylation and expression changes. Chromatin immunoprecipitation followed by qPCR (ChIP-qPCR) and immunofluorescence microscopy provide insights into localized histone modification alterations. Functional assays including cell proliferation measurements, flow cytometry-based cell cycle analysis, and colony formation assays evaluate phenotypic outcomes. Transcriptome-wide RNA-seq further characterizes global gene expression changes. For additional product details or technical support, please contact Ascent Research.
