GATA3 CRISPR Activation Plasmid (m): sc-420495-ACT

Mouse Gata3 encodes the GATA3 transcription factor, a zinc-finger DNA-binding protein that programs lineage specification and differentiation across multiple tissues. In the immune system, GATA3 is a central regulator of T cell development and Th2 polarization, coordinating cytokine gene expression and chromatin remodeling to shape cell fate decisions. It also contributes to epithelial and developmental transcriptional networks by integrating signaling inputs into stable gene expression states. Dysregulated GATA3 activity has been linked to altered immune homeostasis and oncogenic transcriptional programs in cancer-relevant contexts, making it a useful node for mechanistic studies of differentiation, inflammation, and transcriptional control.

GATA3 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Gata3 expression without altering the underlying DNA sequence.

GATA3 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Gata3 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 Gata3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GATA3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Gata3 locus and enabling the study of GATA3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GATA3 pathway restoration in tumor cells with silenced or reduced Gata3 expression.

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