Adenylate cyclase 4/AC4/ADCY4 CRISPR Activation Plasmid (h): sc-403016-ACT

Human ADCY4 encodes adenylate cyclase 4 (AC4), a membrane-associated enzyme that converts ATP to cyclic AMP downstream of G protein–coupled receptor signaling. By modulating cAMP/PKA and EPAC-dependent pathways, AC4 influences phosphorylation cascades, transcriptional programs, and cellular responses such as secretion, metabolism, and inflammatory signaling. ADCY4 activity is integrated with calcium- and Gαs-regulated inputs, shaping the amplitude and duration of second-messenger dynamics across cell types. Altered regulation of cAMP signaling networks involving ADCY4 has been investigated in the context of cardiometabolic traits and immune-related processes, supporting its use as a mechanistic node in pathway studies.

Adenylate cyclase 4/AC4/ADCY4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ADCY4 expression without altering the underlying DNA sequence.

Adenylate cyclase 4/AC4/ADCY4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ADCY4 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 ADCY4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Adenylate cyclase 4/AC4/ADCY4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ADCY4 locus and enabling the study of Adenylate cyclase 4/AC4/ADCY4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Adenylate cyclase 4/AC4/ADCY4 pathway restoration in tumor cells with silenced or reduced ADCY4 expression.

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