ACLP/AEBP1 CRISPR Activation Plasmid (h): sc-403540-ACT

AEBP1 encodes adipocyte enhancer-binding protein 1, also known as ACLP, an extracellular matrix-associated protein implicated in collagen organization, mesenchymal cell differentiation, and regulation of tissue remodeling. In human cells, AEBP1 activity is linked to fibroblast activation programs and crosstalk with TGF-β/SMAD signaling and integrin-mediated adhesion pathways that coordinate ECM deposition and stiffness sensing. Dysregulated AEBP1 expression has been associated with fibrotic phenotypes and stromal remodeling observed across inflammatory microenvironments and tumor-associated stroma. These features make AEBP1 a useful molecular handle for studying matrix biology, cell–matrix signaling, and transcriptional programs governing connective tissue homeostasis.

ACLP/AEBP1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous AEBP1 expression without altering the underlying DNA sequence.

ACLP/AEBP1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the AEBP1 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 AEBP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ACLP/AEBP1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native AEBP1 locus and enabling the study of ACLP/AEBP1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ACLP/AEBP1 pathway restoration in tumor cells with silenced or reduced AEBP1 expression.

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