CART CRISPR Activation Plasmid (h): sc-417375-ACT

Human CARTPT encodes cocaine- and amphetamine-regulated transcript (CART), a neuropeptide precursor processed into bioactive peptides that function as neuromodulators and neuroendocrine signals. CART peptides participate in hypothalamic circuits controlling appetite, energy homeostasis, and reward-related behaviors, integrating endocrine cues with synaptic activity. CARTPT expression is regulated by neuronal activation and metabolic state, linking it to pathways involved in feeding, stress responses, and dopaminergic signaling. Dysregulated CARTPT/CART signaling has been associated with metabolic phenotypes such as obesity and eating behavior traits, as well as neuropsychiatric and addiction-related biology in translational research contexts.

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

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

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