POMC CRISPR Activation Plasmid (h): sc-400592-ACT

Human POMC encodes proopiomelanocortin, a neuroendocrine prohormone that is proteolytically processed into multiple bioactive peptides including ACTH and melanocyte-stimulating hormones. These peptides signal through melanocortin receptors to regulate hypothalamic control of appetite and energy balance, coordinate pituitary–adrenal axis activity, and influence pigmentation and stress responses. POMC expression and peptide processing are tightly linked to cAMP/PKA-driven transcriptional programs and prohormone convertase–dependent maturation in secretory pathways. Dysregulation of POMC pathways is associated with disorders of energy homeostasis, adrenal function, and pigment biology, making it a central target for mechanistic studies of neuroendocrine signaling.

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

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

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