CapG CRISPR Activation Plasmid (h): sc-416526-ACT

CAPG encodes CapG, a calcium-dependent actin-binding protein of the gelsolin/villin family that caps the barbed ends of actin filaments without severing, thereby shaping cortical actin assembly and cytoskeletal remodeling. CapG regulates membrane ruffling, phagocytosis, cell adhesion, and motility, linking extracellular cues to dynamic changes in cell architecture. These activities connect CAPG to pathways governing actin polymerization and cytoskeleton-driven trafficking, including processes coordinated by Rho-family GTPases and integrin-associated signaling. Dysregulated CAPG expression has been associated with altered migratory and invasive phenotypes and is frequently examined in the context of tumor progression, macrophage function, and inflammatory microenvironments.

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

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

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