The CBX2 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HEK293T human embryonic kidney cell line. This product provides a heterogeneous pool of cells with targeted disruption of the CBX2 gene, enabling loss-of-function studies of this Polycomb group repressor. The polyclonal format supports pooled functional assays and high-throughput screens without requiring single-cell cloning.
The host HEK293T cell line is a widely used human embryonic kidney epithelial line transformed with adenovirus 5 DNA and expressing SV40 large T antigen. Known for exceptionally high transfection efficiency and robust viral production, HEK293T is a standard model for molecular biology, protein expression, and lentiviral packaging. Its renal epithelial origin and stable karyotype make it well-suited for epigenetic and signaling studies.
CBX2 (Chromobox 2) is a core subunit of canonical Polycomb Repressive Complex 1 (PRC1). It recognizes trimethylated lysine 27 on histone H3 (H3K27me3) via its chromodomain, a mark deposited by the PRC2 complex containing EZH2. This interaction facilitates chromatin compaction and gene silencing by recruiting RING1A/B E3 ubiquitin ligases, which catalyze monoubiquitination of H2A at lysine 119 (H2AK119ub). CBX2 also interacts with BMI1, PHC1/2/3, and SCMH1 to stabilize PRC1. Its activity is regulated by pluripotency transcription factors SOX2, NANOG, and OCT4, and it represses downstream targets including HOX gene clusters and CDKN2A. Thus, CBX2 serves as a critical link between H3K27me3 and H2AK119ub, mediating stable developmental gene silencing.
In HEK293T cells, CBX2 knockout disrupts a central node of PRC1 function within an epithelial background that endogenously expresses PRC2 core components EZH2, SUZ12, and EED. Loss of CBX2 is expected to impair H2AK119ub deposition and derepress Polycomb target genes, providing a model to dissect PRC1/PRC2 interplay. The presence of SV40 large T antigen, which can influence cell cycle and chromatin modifiers, adds relevance for studying oncogenic chromatin dysregulation.
This knockout pool is applicable to diverse research areas including epigenetic regulation, developmental biology, cancer epigenetics, and sex determination disorders. Researchers can validate CBX2 loss by western blotting and assess H2AK119ub levels, perform RT-qPCR for target gene derepression, and conduct ChIP-qPCR for H3K27me3 occupancy. Co-immunoprecipitation and immunofluorescence enable analysis of PRC1 complex integrity. Functional readouts like proliferation and colony formation assays, along with RNA-seq, can reveal global gene expression changes. For further information, please contact Ascent Research.
