NFATc1 CRISPR Activation Plasmid (m): sc-421863-ACT

Nfatc1 encodes NFATc1, a calcium/calcineurin-regulated transcription factor in the NFAT family that transduces T cell receptor signaling into durable gene expression programs. NFATc1 coordinates immune activation and differentiation by integrating Ca2+ influx with MAPK and AP-1 inputs, shaping cytokine production and lineage decisions in lymphocytes. In mouse, NFATc1 is also implicated in osteoclastogenesis and bone remodeling through RANKL-driven pathways, linking immune signaling to skeletal homeostasis. Dysregulated NFATc1 activity has been associated with inflammatory phenotypes and altered immune cell function, making it a useful node for mechanistic studies of signaling, transcriptional control, and cell fate.

NFATc1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Nfatc1 expression without altering the underlying DNA sequence.

NFATc1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Nfatc1 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 Nfatc1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NFATc1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Nfatc1 locus and enabling the study of NFATc1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NFATc1 pathway restoration in tumor cells with silenced or reduced Nfatc1 expression.

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