CREB1 CRISPR Activation Plasmid (h): sc-400160-ACT

CREB1 (cAMP response element-binding protein 1) is a sequence-specific transcription factor that binds cAMP response elements to coordinate stimulus-dependent gene expression programs. It functions as a central effector of cAMP/PKA signaling and integrates inputs from MAPK/ERK and Ca2+/calmodulin-dependent kinases through phosphorylation-dependent coactivator recruitment, including CBP/p300. CREB1-regulated transcription supports neuronal plasticity and memory formation, circadian and metabolic homeostasis, and pro-survival responses to cellular stress. Dysregulated CREB1 activity and CREB-dependent transcriptional networks have been implicated in oncogenic growth and therapy resistance, neuropsychiatric and neurodegenerative processes, and inflammatory signaling contexts.

CREB1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CREB1 expression without altering the underlying DNA sequence.

CREB1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CREB1 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 CREB1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CREB1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CREB1 locus and enabling the study of CREB1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CREB1 pathway restoration in tumor cells with silenced or reduced CREB1 expression.

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