The BMAL1 Knockout 786-O Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human 786-O renal cell adenocarcinoma cell line, engineered to disrupt the BMAL1 (ARNTL) gene. This loss-of-function model provides a robust tool for examining the roles of the core circadian clock transcription factor BMAL1 in cellular rhythms, metabolic regulation, and cancer biology. The polyclonal nature of the knockout pool ensures representation of diverse edit events, enabling population-level functional studies without clone-specific artifacts.
The 786-O parental line is an adherent epithelial cell line established from a primary clear cell renal cell carcinoma (ccRCC). It harbors a homozygous mutation in the von Hippel?CLindau (VHL) tumor suppressor gene, leading to constitutive stabilization of hypoxia-inducible factors (HIFs) and mimicking the pseudo-hypoxic state characteristic of most ccRCCs. This genetic background makes 786-O a widely used model for investigating VHL/HIF pathway dysregulation, tumor metabolism, angiogenesis, and therapeutic responses in renal cancer.
BMAL1 (Brain and Muscle ARNT-Like 1) is an essential bHLH-PAS transcription factor that heterodimerizes with CLOCK or NPAS2 to bind E-box elements, activating transcription of clock-controlled genes including PER1, PER2, CRY1, CRY2, DBP, and REV-ERB??. Translated PER and CRY proteins multimerize and translocate to the nucleus to repress BMAL1/CLOCK activity, forming a negative feedback loop core to circadian rhythms. Upstream regulation involves REV-ERB?? (repressor) and ROR?? (activator) of BMAL1 transcription, SIRT1-mediated deacetylation, and phosphorylation by casein kinase 1??/??, while SCF-FBXL3 ubiquitin ligase targets CRY for degradation. Through these interactions, BMAL1 coordinates rhythmic expression of metabolic and cell cycle regulators.
In 786-O VHL-deficient renal carcinoma cells, BMAL1 knockout facilitates dissection of crosstalk between circadian rhythms and oncogenic HIF signaling. Circadian disruption is linked to cancer progression, and BMAL1 influences HIF-1?? activity and metabolic reprogramming. Disrupting BMAL1 in this model enables interrogation of how clock loss affects tumor phenotypes such as glycolysis, proliferation under normoxia/hypoxia, and chemosensitivity. The polyclonal population is well-suited for pooled functional screens without clonal bias.
Typical applications include profiling circadian gene expression via RT-qPCR or RNA-seq, evaluating BMAL1 binding dynamics using ChIP-qPCR, assessing cell cycle perturbations by flow cytometry, and performing metabolic flux analyses to link clock disruption to cancer metabolism. Researchers can also employ luciferase reporters of clock gene promoters to study rhythmicity in a cancer background, or combine the knockout with drug sensitivity assays to explore chronotherapeutic strategies. The BMAL1 Knockout 786-O Polyclonal Cells are a versatile platform for translational studies of circadian biology in renal cell carcinoma. For further information or to discuss custom applications, please contact Ascent Research.
