CD13 CRISPR Activation Plasmid (h): sc-401095-ACT

ANPEP encodes CD13 (aminopeptidase N), a zinc-dependent, type II membrane metalloprotease that cleaves N-terminal neutral amino acids from peptides at the cell surface. CD13 participates in peptide metabolism and modulation of cell–cell and cell–matrix interactions, influencing processes such as adhesion, migration, and myeloid lineage biology. Through its enzymatic and non-enzymatic roles, CD13 is implicated in inflammatory signaling networks and remodeling of the extracellular microenvironment. Altered ANPEP/CD13 expression has been associated with hematologic and solid tumor contexts and with immune and vascular pathophysiology, supporting its study as a functional regulator in disease-relevant cell states.

CD13 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ANPEP expression without altering the underlying DNA sequence.

CD13 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ANPEP 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 ANPEP transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CD13 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ANPEP locus and enabling the study of CD13-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CD13 pathway restoration in tumor cells with silenced or reduced ANPEP expression.

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