ECM2 CRISPR Activation Plasmid (m): sc-436571-ACT

Mouse Ecm2 encodes extracellular matrix protein 2 (ECM2), a secreted matrix-associated component implicated in organizing stromal architecture and modulating cell–matrix interactions. ECM2 contributes to extracellular matrix remodeling processes that influence cell adhesion, migration, and tissue morphogenesis, intersecting with pathways that regulate fibroblast activity and basement membrane integrity. Altered ECM composition and ECM2-linked matrix dynamics are relevant to studies of fibrosis-like remodeling, wound repair, and tumor microenvironment biology, where stromal cues shape cellular phenotypes. Ecm2 expression is therefore frequently examined in models of connective tissue homeostasis and inflammation-associated changes in the extracellular milieu.

ECM2 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Ecm2 expression without altering the underlying DNA sequence.

ECM2 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Ecm2 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 Ecm2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ECM2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Ecm2 locus and enabling the study of ECM2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ECM2 pathway restoration in tumor cells with silenced or reduced Ecm2 expression.

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