SGK CRISPR Activation Plasmid (m): sc-422906-ACT

Mouse Sgk1 encodes serum/glucocorticoid regulated kinase (SGK), an AGC family serine/threonine kinase induced by glucocorticoids, growth factors, and cellular stress. SGK integrates PI3K–PDK1 signaling with downstream regulation of ion transporters and channels, transcriptional programs, and survival pathways, including modulation of FOXO-dependent gene expression and cross-talk with mTOR-related nutrient sensing. Through these processes it influences epithelial sodium transport, osmotic and oxidative stress responses, and cell cycle progression. Dysregulated SGK activity has been linked to pathological remodeling and inflammatory signaling contexts, making Sgk1 a useful node for mechanistic studies of stress-adaptation pathways in mammalian tissues.

SGK CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Sgk1 expression without altering the underlying DNA sequence.

SGK CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Sgk1 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 Sgk1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SGK expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Sgk1 locus and enabling the study of SGK-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SGK pathway restoration in tumor cells with silenced or reduced Sgk1 expression.

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