DEC1 CRISPR Activation Plasmid (h): sc-402058-ACT

BHLHE40 encodes the basic helix-loop-helix transcription factor DEC1, a context-dependent regulator of gene expression that integrates environmental cues such as hypoxia and circadian signals. DEC1 modulates transcriptional programs linked to cell-cycle control, differentiation, epithelial–mesenchymal transition, and metabolic adaptation, and it interfaces with pathways including HIF-1 signaling and core clock networks. Through these functions, DEC1 contributes to regulation of stress responses and inflammatory gene expression, with reported associations to tumor biology, fibrosis-related remodeling, and immune dysregulation in diverse experimental systems.

DEC1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous BHLHE40 expression without altering the underlying DNA sequence.

DEC1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the BHLHE40 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 BHLHE40 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DEC1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native BHLHE40 locus and enabling the study of DEC1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DEC1 pathway restoration in tumor cells with silenced or reduced BHLHE40 expression.

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