ATF3 CRISPR Activation Plasmid (m): sc-419227-ACT

Mouse Atf3 encodes activating transcription factor 3 (ATF3), a stress-inducible bZIP transcription factor that integrates signals from MAPK/JNK, TLR/NF-κB, and integrated stress response pathways to reshape context-dependent gene expression programs. ATF3 modulates inflammatory transcriptional circuits, apoptosis and survival decisions, and metabolic adaptation by binding CRE/ATF elements and coordinating with other AP-1 family factors. In immune and stromal compartments, ATF3 acts as a feedback regulator of cytokine and chemokine expression, influencing macrophage activation states and tissue injury responses. Dysregulated ATF3 activity has been linked to models of neuroinflammation, metabolic stress, and tumor-associated signaling, making it useful for probing transcriptional control of stress and innate immune phenotypes.

ATF3 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Atf3 expression without altering the underlying DNA sequence.

ATF3 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Atf3 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 Atf3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ATF3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Atf3 locus and enabling the study of ATF3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ATF3 pathway restoration in tumor cells with silenced or reduced Atf3 expression.

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