ANG I CRISPR Activation Plasmid (h): sc-401766-ACT

Human ANG encodes angiogenin, a secreted ribonuclease that binds cell-surface receptors and extracellular matrix components to influence endothelial cell behavior, stress responses, and RNA metabolism. ANG participates in processes linked to angiogenesis and nutrient signaling, including modulation of rRNA/tRNA-derived fragments and downstream effects on protein synthesis and cell survival. Dysregulated ANG expression or activity has been associated with vascular remodeling and neurodegenerative phenotypes, and is frequently examined in contexts of inflammation, hypoxia, and tumor microenvironment biology. These functions make ANG a useful entry point for studying paracrine signaling, RNA cleavage–dependent stress programs, and pathway crosstalk that shapes tissue homeostasis.

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

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

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