Endo180 CRISPR Activation Plasmid (h): sc-403483-ACT

MRC2 encodes Endo180 (uPARAP), a transmembrane endocytic receptor of the mannose receptor family that binds and internalizes collagens and other extracellular matrix ligands for lysosomal degradation. Endo180 activity supports extracellular matrix turnover, cell adhesion and migration, and remodeling of the tumor microenvironment through collagen uptake and trafficking pathways coupled to endosomal–lysosomal processing. The receptor is frequently studied in fibroblasts, macrophage-lineage cells, and invasive carcinoma models where collagen dynamics shape tissue architecture and signaling. Dysregulated MRC2/Endo180 expression is associated with fibrotic remodeling and cancer invasion biology, making it relevant for mechanistic studies of matrix-driven cell behavior.

Endo180 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MRC2 expression without altering the underlying DNA sequence.

Endo180 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MRC2 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 MRC2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Endo180 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MRC2 locus and enabling the study of Endo180-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Endo180 pathway restoration in tumor cells with silenced or reduced MRC2 expression.

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