ATF-2 CRISPR Activation Plasmid (h): sc-416662-ACT

Human ATF2 encodes activating transcription factor 2 (ATF-2), a bZIP transcription factor activated by stress-responsive kinases including JNK and p38 MAPK. Upon phosphorylation, ATF-2 regulates inducible gene programs controlling cell-cycle progression, DNA damage responses, inflammation, and apoptosis, often through heterodimerization with AP-1 family members and integration of MAPK signaling. ATF-2 activity intersects with chromatin regulation and immediate-early transcription, influencing context-specific programs in immune signaling and cellular adaptation to genotoxic and oxidative stress. Dysregulated ATF-2 signaling has been associated with altered proliferative and stress-response phenotypes reported across diverse cancer and inflammatory disease models, supporting its use as a mechanistic node in pathway studies.

ATF-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ATF2 expression without altering the underlying DNA sequence.

ATF-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ATF2 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 ATF2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ATF-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ATF2 locus and enabling the study of ATF-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ATF-2 pathway restoration in tumor cells with silenced or reduced ATF2 expression.

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