Acrp30 CRISPR Activation Plasmid (h): sc-401327-ACT

ADIPOQ encodes the adipokine Acrp30 (adiponectin), a secreted protein primarily produced by adipocytes that circulates as multimeric complexes and regulates systemic energy homeostasis. Acrp30 engages receptors such as ADIPOR1/ADIPOR2 to modulate AMPK and PPARα signaling, promoting fatty-acid oxidation, glucose utilization, and anti-inflammatory programs in metabolic tissues. Altered ADIPOQ expression and adiponectin signaling are linked to insulin resistance, obesity-associated metabolic dysfunction, nonalcoholic fatty liver disease, and cardiovascular risk phenotypes. In cell models, ADIPOQ is used to interrogate adipocyte differentiation, adipokine secretion, and crosstalk between adipose tissue and liver, muscle, and immune pathways.

Acrp30 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ADIPOQ expression without altering the underlying DNA sequence.

Acrp30 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ADIPOQ 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 ADIPOQ transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Acrp30 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ADIPOQ locus and enabling the study of Acrp30-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Acrp30 pathway restoration in tumor cells with silenced or reduced ADIPOQ expression.

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