Per2 CRISPR Activation Plasmid (h): sc-401089-ACT

Human PER2 encodes the core circadian regulator Per2, a PAS domain–containing protein that participates in the transcriptional–translational feedback loops controlling 24-hour rhythmicity. PER2 coordinates CLOCK/ARNTL (BMAL1)-driven transcription by modulating repressor complex dynamics and integrates timing information with cellular programs such as metabolism, DNA damage responses, and cell-cycle progression. Through coupling circadian transcription to signaling pathways including glucocorticoid, AMPK, and MAPK-linked networks, PER2 influences tissue-specific rhythmic gene expression. Dysregulated PER2 activity and circadian misalignment have been associated with sleep–wake disorders and altered susceptibility to oncogenic and metabolic phenotypes, making it a useful target in systems biology and mechanistic studies.

Per2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PER2 expression without altering the underlying DNA sequence.

Per2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PER2 locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the PER2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Per2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PER2 locus and enabling the study of Per2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Per2 pathway restoration in tumor cells with silenced or reduced PER2 expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.