FAPα CRISPR Activation Plasmid (h): sc-401511-ACT

Fibroblast activation protein alpha (FAPα) is a membrane-anchored serine protease of the dipeptidyl peptidase family that exhibits both dipeptidyl peptidase and endopeptidase activities, contributing to extracellular matrix remodeling. FAP is typically enriched in activated fibroblasts and stromal compartments, where it can influence cell–matrix interactions, tissue remodeling programs, and paracrine signaling within the microenvironment. Its activity has been linked to fibroblast activation states, collagen turnover, and regulation of proteolytic networks that intersect with wound-healing and fibrotic processes. Dysregulated FAP expression is frequently studied in contexts involving stromal remodeling and inflammation-associated microenvironment changes, supporting research into fibrosis biology and tumor-associated stroma.

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

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

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