ATF-6α CRISPR Activation Plasmid (h): sc-400259-ACT

ATF6 encodes activating transcription factor 6α, an ER membrane–tethered bZIP transcription factor that functions as a primary sensor and effector of the unfolded protein response (UPR). Upon ER stress, ATF-6α is transported to the Golgi for regulated intramembrane proteolysis, releasing a nuclear fragment that induces genes involved in ER chaperoning, ER-associated degradation (ERAD), and proteostasis. Through cross-talk with PERK–eIF2α–ATF4 and IRE1–XBP1 signaling, ATF-6α coordinates adaptive remodeling of secretory pathway capacity and cellular stress resilience. Dysregulated ATF6/UPR signaling is implicated in contexts such as metabolic dysfunction, inflammation, neurodegeneration, and tumor cell stress adaptation, making ATF6 a useful node for mechanistic studies of proteostasis and stress signaling.

ATF-6α CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ATF6 expression without altering the underlying DNA sequence.

ATF-6α CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ATF6 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 ATF6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ATF-6α expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ATF6 locus and enabling the study of ATF-6α-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ATF-6α pathway restoration in tumor cells with silenced or reduced ATF6 expression.

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