GILZ CRISPR Activation Plasmid (h): sc-401424-ACT

TSC22D3 encodes glucocorticoid-induced leucine zipper (GILZ), a stress- and hormone-responsive regulator that links glucocorticoid receptor signaling to transcriptional control of immune and inflammatory programs. GILZ modulates pathways including NF-κB and AP-1, shaping cytokine expression, T cell activation, and macrophage polarization, and it can influence cell survival and differentiation through interactions with MAPK and PI3K/AKT-related signaling nodes. Altered TSC22D3/GILZ expression has been associated with dysregulated inflammatory states and immune-mediated pathology, and it is frequently studied in contexts where glucocorticoid responsiveness, tolerance, or immune suppression mechanisms are relevant. These properties make GILZ a useful molecular handle for dissecting transcriptional networks that govern inflammation, stress adaptation, and lineage-specific immune functions in human cells.

GILZ CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TSC22D3 expression without altering the underlying DNA sequence.

GILZ CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TSC22D3 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 TSC22D3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GILZ expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TSC22D3 locus and enabling the study of GILZ-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GILZ pathway restoration in tumor cells with silenced or reduced TSC22D3 expression.

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