CRF-BP CRISPR Activation Plasmid (h): sc-403849-ACT

CRHBP encodes corticotropin-releasing factor–binding protein (CRF-BP), a secreted glycoprotein that binds CRF-family peptides and modulates their bioavailability, receptor engagement, and peptide clearance. By shaping CRF signaling, CRF-BP influences hypothalamic–pituitary–adrenal (HPA) axis regulation, neuroendocrine stress responses, and downstream cAMP/PKA-dependent transcriptional programs. CRHBP expression and CRF/urocortin pathway activity have been implicated in stress-related neuropsychiatric phenotypes, inflammatory signaling, and metabolic homeostasis. In experimental systems, CRF-BP provides a mechanistic handle to study ligand buffering and context-dependent CRF receptor signaling in neuronal and peripheral cell types.

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

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

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