Adenylate cyclase 2/AC2/ADCY2 CRISPR Activation Plasmid (h): sc-402994-ACT

Human ADCY2 encodes adenylate cyclase 2 (AC2), a membrane-bound enzyme that converts ATP to cyclic AMP (cAMP) in response to G protein–coupled receptor signaling, particularly via stimulatory Gαs and modulatory Gβγ inputs. By tuning intracellular cAMP, AC2 regulates PKA- and EPAC-dependent phosphorylation programs that influence ion channel activity, neurotransmitter release, metabolic responses, and transcriptional outputs such as CREB signaling. ADCY2 activity is therefore integral to signal integration at the plasma membrane and to the temporal dynamics of second-messenger propagation in excitable and non-excitable cells. Genetic and functional studies have linked altered cAMP pathway regulation involving ADCY2 to neuropsychiatric and behavioral phenotypes, supporting its relevance for mechanistic studies of neuronal signaling and cellular homeostasis.

Adenylate cyclase 2/AC2/ADCY2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ADCY2 expression without altering the underlying DNA sequence.

Adenylate cyclase 2/AC2/ADCY2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ADCY2 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 ADCY2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Adenylate cyclase 2/AC2/ADCY2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ADCY2 locus and enabling the study of Adenylate cyclase 2/AC2/ADCY2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Adenylate cyclase 2/AC2/ADCY2 pathway restoration in tumor cells with silenced or reduced ADCY2 expression.

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