Annexin-2/ANXA2 CRISPR Activation Plasmid (h): sc-400296-ACT

Human ANXA2 encodes annexin A2, a Ca²⁺-dependent phospholipid-binding protein that assembles with S100A10 to regulate membrane organization, endocytosis, and actin cytoskeletal dynamics. Annexin A2 coordinates plasmin generation and fibrinolytic activity at the cell surface, influencing extracellular matrix remodeling, cell migration, and angiogenic processes. It participates in vesicle trafficking and signaling events linked to inflammatory responses and oxidative stress, and can modulate plasminogen activation and protease localization. Dysregulated ANXA2 expression or localization has been reported across multiple cancer types and vascular and inflammatory pathologies, supporting its use as a mechanistic node in studies of invasion, metastasis-associated programs, and thrombosis-related biology.

Annexin-2/ANXA2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ANXA2 expression without altering the underlying DNA sequence.

Annexin-2/ANXA2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ANXA2 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 ANXA2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Annexin-2/ANXA2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ANXA2 locus and enabling the study of Annexin-2/ANXA2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Annexin-2/ANXA2 pathway restoration in tumor cells with silenced or reduced ANXA2 expression.

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