CRIM1 CRISPR Activation Plasmid (h): sc-404886-ACT

CRIM1 (cysteine rich transmembrane BMP regulator 1) encodes a transmembrane protein that modulates extracellular signaling by binding and presenting growth factors, particularly within BMP/TGF-β networks. It contributes to developmental patterning, cell–cell and cell–matrix interactions, and regulation of endothelial and epithelial behavior through control of morphogen availability and receptor engagement. CRIM1 activity has been linked to processes including angiogenesis, organ morphogenesis, and tissue homeostasis, where altered expression can shift differentiation and remodeling programs. Dysregulation of CRIM1 has been reported in contexts of developmental abnormalities and cancer-associated phenotypes, supporting its use as a node to study growth factor signaling balance and microenvironmental control.

CRIM1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CRIM1 expression without altering the underlying DNA sequence.

CRIM1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CRIM1 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 CRIM1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CRIM1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CRIM1 locus and enabling the study of CRIM1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CRIM1 pathway restoration in tumor cells with silenced or reduced CRIM1 expression.

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