gremlin-2 CRISPR Activation Plasmid (h): sc-403982-ACT

Human GREM2 encodes gremlin-2, a secreted cystine-knot glycoprotein that functions as an extracellular antagonist of bone morphogenetic proteins (BMPs), including BMP2 and BMP4. By sequestering BMP ligands, gremlin-2 modulates SMAD-dependent transcriptional programs that shape cell fate decisions, differentiation, and tissue patterning, and interfaces with broader TGF-β superfamily signaling. GREM2 activity is therefore relevant to studies of developmental biology, fibrosis-associated remodeling, and regulation of osteogenic and neurovascular processes. Dysregulated BMP antagonism has been linked in the literature to disease-associated changes in extracellular matrix dynamics and aberrant differentiation states, making GREM2 a useful target for mechanistic pathway interrogation.

gremlin-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GREM2 expression without altering the underlying DNA sequence.

gremlin-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GREM2 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 GREM2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous gremlin-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GREM2 locus and enabling the study of gremlin-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of gremlin-2 pathway restoration in tumor cells with silenced or reduced GREM2 expression.

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