GATA4 CRISPR Activation Plasmid (h): sc-400122-ACT

GATA4 encodes a zinc-finger transcription factor that binds GATA motifs to regulate gene programs controlling cardiac morphogenesis, cardiomyocyte differentiation, and endoderm-derived tissue development. It functions within lineage-specifying networks alongside factors such as NKX2-5 and TBX5, integrating signaling inputs to modulate chromatin accessibility and transcriptional output during development and stress responses. In adult tissues, GATA4 influences cardiac hypertrophic signaling and survival pathways, and altered expression or variants have been associated with congenital heart defects and cardiomyopathies. Dysregulated GATA4 activity is also studied in contexts of gastrointestinal differentiation and oncogenic transcriptional reprogramming.

GATA4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GATA4 expression without altering the underlying DNA sequence.

GATA4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GATA4 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 GATA4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GATA4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GATA4 locus and enabling the study of GATA4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GATA4 pathway restoration in tumor cells with silenced or reduced GATA4 expression.

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