The BMAL2 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited heterogeneous cell population with targeted disruption of the circadian transcription factor BMAL2 (ARNTL2). This polyclonal knockout model consists of a pool of HEK293T cells carrying diverse loss-of-function mutations, generated by introducing guide RNAs and Cas9 nuclease, which collectively ablate BMAL2 protein expression. The polyclonal format minimizes clonal bias and provides a robust system for studying circadian transcriptional regulation.
HEK293T cells derive from human embryonic kidney epithelium and are immortalized via stable expression of SV40 large T antigen, which enhances transfection efficiency and recombinant protein production. Though not endogenously rhythmic, these cells express core clock components and can be synchronized with serum shock or chemical agents, allowing reconstitution of circadian oscillations. This tractable and scalable cell line supports high-resolution biochemical and genetic analyses of clock mechanisms.
BMAL2 is a basic helix-loop-helix-PAS transcription factor that heterodimerizes with CLOCK or NPAS2 to bind E-box elements (CACGTG) and activate target gene transcription. Direct targets include the negative feedback genes PER1, PER2, CRY1, and CRY2, as well as output genes DBP and REV-ERB??. Accumulated PER/CRY complexes translocate into the nucleus and inhibit BMAL2-CLOCK/NPAS2 activity, forming the core circadian negative feedback loop. BMAL2 function is modulated by upstream regulators such as SIRT1 deacetylase, AMPK, CK1??, and GSK3?? kinases, and its stability is influenced by partners CIPC and FBXL3. An interlocking REV-ERB??/ROR?? loop further regulates BMAL2 expression, connecting circadian timing to metabolic and stress signaling pathways.
Knocking out BMAL2 in HEK293T eliminates the positive limb of the circadian oscillator, disabling E-box-driven transcription of clock-controlled genes and providing a clean loss-of-function background for mechanistic rescue experiments. The high transfectability of HEK293T cells facilitates episomal or stable re-introduction of BMAL2 variants, enabling structure-function studies and drug screening. The polyclonal nature avoids clonal artifacts, ensuring that phenotypes are attributable to gene disruption rather than cellular drift, and supports robust, reproducible data across a diverse genetic background.
This knockout product is suited for circadian rhythm research, chronopharmacology, metabolic syndrome, and cancer chronotherapy studies. Researchers can validate knockout via Sanger sequencing and assess downstream effects using RT-qPCR and western blotting for PER and CRY proteins. Functional analysis includes circadian luciferase reporter assays to measure oscillatory dynamics, co-immunoprecipitation for protein complexes, ChIP-qPCR for promoter occupancy, and metabolic flux analysis to probe clock-metabolism crosstalk. For further technical details or ordering information, please contact Ascent Research.
