MFAP4 CRISPR Activation Plasmid (h): sc-405012-ACT

MFAP4 (microfibril-associated glycoprotein 4) encodes an extracellular matrix glycoprotein enriched in elastic fiber–associated microfibrils and expressed by mesenchymal and vascular cell types. MFAP4 participates in matrix organization and cell–matrix communication, supporting adhesion and signaling through integrin-linked pathways that influence smooth muscle cell behavior, fibroblast activity, and tissue remodeling. Dysregulated MFAP4 expression and deposition are frequently studied in contexts of vascular remodeling, fibrosis-associated extracellular matrix turnover, and inflammatory stromal responses. As a matrix-associated factor, MFAP4 serves as a useful readout and modulator in models examining extracellular matrix dynamics, mechanotransduction, and microenvironment-dependent phenotypes.

MFAP4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MFAP4 expression without altering the underlying DNA sequence.

MFAP4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MFAP4 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 MFAP4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MFAP4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MFAP4 locus and enabling the study of MFAP4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MFAP4 pathway restoration in tumor cells with silenced or reduced MFAP4 expression.

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