chordin CRISPR Activation Plasmid (h): sc-404997-ACT

Human CHRD encodes chordin, a secreted cysteine-rich glycoprotein that antagonizes BMP signaling by binding BMP ligands in the extracellular space and limiting their receptor engagement. By shaping BMP/TGF-β pathway activity, chordin helps regulate embryonic patterning, mesodermal specification, and tissue morphogenesis, with downstream impacts on SMAD-mediated transcriptional programs. Dysregulated CHRD expression or altered BMP antagonism has been linked to developmental abnormalities and disease-associated remodeling processes where BMP gradients influence cell fate, differentiation, and extracellular matrix organization. CHRD is therefore relevant for studies of morphogen signaling dynamics, lineage commitment, and context-dependent BMP pathway modulation in human cells.

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

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

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