GADD 45γ CRISPR Activation Plasmid (h): sc-402483-ACT

GADD45G (GADD45γ) is a stress-responsive member of the GADD45 family that links genotoxic and inflammatory cues to control of cell-cycle checkpoints, DNA repair signaling, and apoptosis. It participates in pathways centered on MAPK signaling, p53-dependent stress responses, and modulation of CDK activity to enforce growth arrest during cellular damage or differentiation programs. Through these functions, GADD45γ contributes to maintenance of genomic stability and regulation of immune and hematopoietic cell states. Altered GADD45G expression has been associated with dysregulated proliferation, aberrant stress signaling, and epigenetic silencing patterns reported across multiple malignancy contexts, making it a useful node for mechanistic studies of tumor suppressive networks.

GADD 45γ CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GADD45G expression without altering the underlying DNA sequence.

GADD 45γ CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GADD45G 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 GADD45G transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GADD 45γ expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GADD45G locus and enabling the study of GADD 45γ-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GADD 45γ pathway restoration in tumor cells with silenced or reduced GADD45G expression.

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