GDF-15 CRISPR Activation Plasmid (h): sc-401430-ACT

Growth differentiation factor 15 (GDF15) encodes GDF-15, a stress-responsive secreted cytokine in the TGF-β superfamily that is induced by mitochondrial dysfunction, hypoxia, and inflammatory cues. GDF-15 influences cellular adaptation programs including integrated stress responses, metabolic remodeling, and regulation of immune cell activity, with downstream effects on apoptosis, proliferation, and tissue repair. Altered GDF-15 expression has been associated with cardiometabolic disorders, cancer-associated stress signaling, neuroinflammation, and systemic inflammatory states, making it a useful node for pathway interrogation. As a circulating factor, it is also leveraged to study intercellular communication between diseased tissues and immune or stromal compartments.

GDF-15 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GDF15 expression without altering the underlying DNA sequence.

GDF-15 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GDF15 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 GDF15 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GDF-15 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GDF15 locus and enabling the study of GDF-15-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GDF-15 pathway restoration in tumor cells with silenced or reduced GDF15 expression.

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