ATE1 CRISPR Activation Plasmid (h): sc-405539-ACT

ATE1 encodes arginyltransferase 1, a key enzyme in the N-degron (N-end rule) pathway that catalyzes post-translational arginylation of proteins to generate degrons recognized by ubiquitin ligases. Through regulation of ubiquitin–proteasome–mediated turnover, ATE1 influences protein homeostasis, cytoskeletal organization, cell migration, and stress-adaptive signaling. ATE1-dependent arginylation has also been linked to quality control of misfolded proteins and modulation of proteostasis networks. Dysregulated N-degron signaling and altered protein degradation dynamics involving ATE1 are studied in contexts such as cancer cell behavior and neurodegeneration-associated proteotoxic stress.

ATE1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ATE1 expression without altering the underlying DNA sequence.

ATE1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ATE1 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 ATE1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ATE1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ATE1 locus and enabling the study of ATE1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ATE1 pathway restoration in tumor cells with silenced or reduced ATE1 expression.

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