GSK3 beta CRISPR Activation Plasmid (m): sc-425249-ACT

Mouse Gsk3b encodes glycogen synthase kinase 3 beta (GSK3 beta), a serine/threonine kinase that integrates signals from PI3K–AKT, Wnt/β-catenin, Hedgehog, and inflammatory pathways to control phosphorylation-dependent protein turnover and transcriptional programs. GSK3 beta regulates glycogen metabolism, cell cycle progression, apoptosis, cytoskeletal dynamics, and neuronal development through substrates that include β-catenin and multiple transcription factors. Dysregulated GSK3 beta activity has been linked to altered insulin signaling and metabolic homeostasis, aberrant Wnt signaling, and changes in synaptic plasticity and neuroinflammatory states. In mouse models, modulation of Gsk3b expression is commonly used to interrogate pathway crosstalk affecting proliferation, differentiation, and stress responses in diverse tissues.

GSK3 beta CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Gsk3b expression without altering the underlying DNA sequence.

GSK3 beta CRISPR Activation Plasmid (m) 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.