AGRP CRISPR Activation Plasmid (h): sc-402410-ACT

Human AGRP (agouti-related neuropeptide) encodes an orexigenic neuropeptide predominantly produced by hypothalamic neurons that regulate energy homeostasis and feeding behavior. AGRP functions as an inverse agonist/antagonist of melanocortin receptors, especially MC3R and MC4R, opposing α-MSH signaling to modulate cAMP-dependent neuroendocrine pathways controlling appetite, thermogenesis, and glucose metabolism. Through integration with leptin, ghrelin, and insulin-responsive neuronal circuits, AGRP contributes to nutrient-sensing and adaptive responses to fasting. Dysregulated AGRP signaling has been linked to obesity-related phenotypes, metabolic imbalance, and altered neuroendocrine control of body weight.

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

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

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