BMI1 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the BMI1 gene in the human HEK293T cell line. This polyclonal format provides a genetically diverse pool of cells carrying targeted gene disruptions, enabling robust functional studies of BMI1 loss without the confounding effects of clonal selection. The population-level knockout ensures representation of various editing events, making it ideal for assays requiring average population behavior.
HEK293T cells are human embryonic kidney epithelial cells transformed with adenovirus 5 DNA and stably expressing SV40 large T antigen. This transformation endows them with high proliferative capacity, ease of transfection, and partial inactivation of p53 and Rb tumor suppressors. These characteristics make HEK293T a standard host for studying gene function, particularly for genes involved in cell cycle control and senescence, where the SV40 background can help isolate specific pathways.
BMI1 encodes a transcriptional repressor that serves as a core component of PRC1, critical for chromatin remodeling and gene silencing. BMI1 directly represses the INK4a/ARF locus, suppressing p16INK4a and p14ARF expression. Its activity is regulated by upstream signals including c-Myc, E2F, and Sonic Hedgehog. BMI1 partners with RING1B, CBX family members, and PRC2 components like EZH2 to deposit H3K27me3 marks. Disruption of BMI1 removes PRC1-mediated repression, leading to upregulation of p16INK4a and p14ARF, which can trigger cell cycle arrest, senescence, or apoptosis.
In the HEK293T context, BMI1 knockout creates a model to examine the interplay between PRC1 and cell cycle regulation. Despite SV40 large T-mediated inactivation of p53 and Rb, BMI1 loss still elevates p16INK4a, which can inhibit CDK4/6 and restore Rb function, inducing senescence-like phenotypes. This system permits dissection of BMI1-dependent and -independent pathways governing proliferation and senescence, and serves as a platform for studying PRC1/PRC2 crosstalk and histone modification dynamics without interference from functional p53.
These polyclonal knockout cells are suitable for epigenetic regulation studies, cancer research (including glioblastoma, leukemia, prostate, breast, and medulloblastoma), cell cycle and senescence assays, and drug screening for BMI1 inhibitors. Standard assays include Western blotting for BMI1, p16INK4a, and p14ARF; RT-qPCR; ChIP-seq for H3K27me3; proliferation assays (MTS, BrdU); senescence-associated ??-galactosidase staining; flow cytometry for cell cycle and apoptosis; and migration/invasion assays. For further technical details, please contact Ascent Research.
