Noggin CRISPR Activation Plasmid (h): sc-402090-ACT

Human NOG encodes noggin, a secreted cystine-knot glycoprotein that functions as a high-affinity antagonist of BMP family ligands, including BMP2, BMP4, and BMP7, thereby constraining SMAD1/5/8 signaling. By sequestering BMPs, noggin helps regulate embryonic patterning, osteo-chondrogenic differentiation, and tissue morphogenesis, with downstream effects on cell fate decisions and extracellular matrix programs. Altered NOG dosage or activity is associated with developmental skeletal phenotypes and congenital malformations involving joint formation and craniofacial or limb patterning, reflecting dysregulated BMP pathway output. In vitro, NOG serves as a key node for dissecting BMP/TGF-β pathway crosstalk during differentiation and morphogen-driven signaling gradients.

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

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

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