Akt3 CRISPR Activation Plasmid (h): sc-400028-ACT

AKT3 encodes the serine/threonine kinase Akt3, a PI3K-dependent effector that integrates growth factor and nutrient cues to regulate cell survival, metabolism, proliferation, and differentiation. Akt3 participates in canonical PI3K–AKT–mTOR signaling and intersects with FOXO, GSK3, and BAD-mediated networks that shape apoptosis resistance, protein synthesis, and cell cycle progression. In human tissues, AKT3 activity is particularly relevant to neural development and cellular stress responses, and dysregulated signaling has been associated with altered growth control and oncogenic pathway remodeling across multiple disease contexts. Modulating AKT3 expression is therefore useful for dissecting pathway dynamics, feedback regulation, and cell-state transitions linked to proliferative and metabolic phenotypes.

Akt3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous AKT3 expression without altering the underlying DNA sequence.

Akt3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the AKT3 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 AKT3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Akt3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native AKT3 locus and enabling the study of Akt3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Akt3 pathway restoration in tumor cells with silenced or reduced AKT3 expression.

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