CRY1 CRISPR Activation Plasmid (h): sc-400946-ACT

Human CRY1 encodes cryptochrome circadian regulator 1, a core component of the molecular clock that forms repressive complexes with PER proteins to inhibit CLOCK:BMAL1-driven transcription. Through transcriptional–translational feedback loops, CRY1 helps coordinate rhythmic expression programs that influence cell cycle progression, DNA damage responses, metabolism, and endocrine signaling. Dysregulated CRY1 activity has been linked to altered circadian phase and sleep–wake timing, and circadian disruption is frequently associated with metabolic and neurobehavioral phenotypes. In biomedical research, CRY1 is commonly studied to interrogate clock-controlled gene networks and time-of-day effects on cellular physiology.

CRY1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CRY1 expression without altering the underlying DNA sequence.

CRY1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CRY1 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 CRY1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CRY1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CRY1 locus and enabling the study of CRY1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CRY1 pathway restoration in tumor cells with silenced or reduced CRY1 expression.

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