DEC2 CRISPR Activation Plasmid (h): sc-402891-ACT

BHLHE41 encodes the basic helix-loop-helix transcription factor DEC2, a nuclear regulator that binds E-box motifs to modulate circadian transcriptional networks and cell state programs. DEC2 integrates inputs from core clock components and couples rhythmic gene expression to processes such as sleep–wake timing, metabolic adaptation, and cellular stress responses, including hypoxia-associated transcriptional remodeling. In immune and epithelial contexts, DEC2 has been linked to differentiation and inflammatory signaling by shaping transcriptional outputs downstream of environmental cues. Dysregulated BHLHE41/DEC2 activity has been associated in the literature with altered sleep phenotypes and with context-dependent roles in tumor biology and metabolic disease models, supporting mechanistic studies of transcriptional control.

DEC2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous BHLHE41 expression without altering the underlying DNA sequence.

DEC2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the BHLHE41 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 BHLHE41 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DEC2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native BHLHE41 locus and enabling the study of DEC2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DEC2 pathway restoration in tumor cells with silenced or reduced BHLHE41 expression.

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