Description

The PER2 Knockout H4 Cell Line is a CRISPR/Cas9-edited knockout cell line based on H4 human neuroglioma cells. It features a targeted disruption of PER2, generating a stable loss-of-function model for circadian clock studies. This CRISPR/Cas9-mediated gene disruption eliminates PER2 protein expression, enabling functional dissection without off-target effects of transient methods. The line is optimized for complementation assays and signaling pathway analysis.

The H4 cell line was established from a human brain glioma and retains glial cell characteristics, including support and insulation functions for neurons. H4 cells are widely used for glial biology and brain tumor research due partly to their robust growth and genetic manipulability, making them ideal for CRISPR-based circadian gene knockout.

PER2 acts as a transcriptional repressor in the circadian negative feedback loop. It is transcriptionally induced by CLOCK-BMAL1 heterodimers, phosphorylated by CK1??/?? and GSK3??, and then forms complexes with CRY1/CRY2. These PER-CRY complexes translocate to the nucleus to inhibit CLOCK-BMAL1-mediated transcription of clock genes (PER1, CRY, DBP) and cell cycle regulators. Additional interacting factors include TIMELESS and CK1??/??, linking PER2 to diverse physiological rhythms.

The H4 neuroglioma background makes this line particularly valuable for dissecting glial circadian biology. Astrocytic clocks regulate synaptic activity, metabolic coupling, and neuroinflammation. PER2 disruption in H4 cells thus enables exploration of glial clock dysfunction in glioma progression, sleep disorders, and metabolic imbalances. Moreover, glial circadian disruption may influence drug metabolism and chronotherapeutic outcomes, expanding application to neuropharmacology.

This knockout line supports diverse assays: real-time bioluminescent recording of circadian dynamics via luciferase reporters, RT-qPCR profiling of rhythmic gene expression, western blotting for PER2 and phosphorylation variants, and immunofluorescence for subcellular localization. Co-immunoprecipitation can be performed to study PER2-CRY1/2 complexes, and flow cytometry enables cell cycle analysis. Drug sensitivity testing facilitates chronotherapeutic investigations, and overexpressing mutant PER2 models sleep phase syndromes. For further information, please contact Ascent Research.