NFATc3 CRISPR Activation Plasmid (h): sc-400339-ACT

NFATC3 encodes NFATc3, a calcium/calcineurin-regulated transcription factor that translocates to the nucleus to coordinate stimulus-dependent gene expression programs. NFATc3 integrates signaling from T cell receptor and other immunoreceptor pathways with MAPK and AP-1 inputs to regulate cytokine production, differentiation, and cellular activation states. Beyond immune lineages, NFATc3 contributes to transcriptional control in cardiovascular and neural contexts through calcium-dependent transcriptional networks. Dysregulated NFAT signaling has been associated with inflammatory pathophysiology and cancer-related phenotypes, making NFATC3 a useful node for mechanistic studies of signal-to-transcription coupling.

NFATc3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NFATC3 expression without altering the underlying DNA sequence.

NFATc3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NFATC3 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 NFATC3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NFATc3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NFATC3 locus and enabling the study of NFATc3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NFATc3 pathway restoration in tumor cells with silenced or reduced NFATC3 expression.

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