ATF4 CRISPR Activation Plasmid (m): sc-419228-ACT

Mouse Atf4 encodes activating transcription factor 4 (ATF4), a stress-responsive bZIP transcription factor that integrates signals from the integrated stress response and unfolded protein response to reshape gene expression programs. ATF4 is induced downstream of eIF2α phosphorylation and coordinates amino acid metabolism, redox and glutathione homeostasis, autophagy, and adaptive survival pathways during nutrient limitation, ER stress, and hypoxia. Through crosstalk with PERK–eIF2α, mTOR, and oxidative stress signaling, ATF4 influences cellular differentiation and mitochondrial function in multiple tissues. Dysregulated ATF4 activity has been associated with pathological stress adaptation and inflammatory and metabolic phenotypes, making it a useful node for mechanistic studies of proteostasis and stress-linked disease models in mice.

ATF4 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Atf4 expression without altering the underlying DNA sequence.

ATF4 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Atf4 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 Atf4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ATF4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Atf4 locus and enabling the study of ATF4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ATF4 pathway restoration in tumor cells with silenced or reduced Atf4 expression.

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