GSK3 beta CRISPR Activation Plasmid (h): sc-400090-ACT

Human GSK3B encodes glycogen synthase kinase 3 beta (GSK3β), a constitutively active serine/threonine kinase that integrates nutrient and growth factor cues to coordinate cell fate decisions. GSK3β is a central negative regulator of canonical Wnt/β-catenin signaling and also participates in PI3K–AKT, insulin/glycogen metabolism, Hedgehog, Notch, and NF-κB-associated processes, shaping transcription, proliferation, apoptosis, and differentiation. Through phosphorylation of diverse substrates such as β-catenin and multiple transcriptional regulators, GSK3β influences cytoskeletal dynamics, cell cycle control, and metabolic homeostasis. Dysregulated GSK3B signaling has been implicated across research contexts including cancer biology, neurodegeneration-associated pathways, inflammation, and metabolic disease models.

GSK3 beta CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GSK3B expression without altering the underlying DNA sequence.

GSK3 beta CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GSK3B 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 GSK3B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GSK3 beta expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GSK3B locus and enabling the study of GSK3 beta-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GSK3 beta pathway restoration in tumor cells with silenced or reduced GSK3B expression.

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