Prokineticin-2 CRISPR Activation Plasmid (h): sc-403930-ACT

Human PROK2 encodes prokineticin-2, a secreted chemokine-like peptide that signals primarily through the G protein-coupled receptors PROKR1 and PROKR2 to regulate calcium flux, MAPK/ERK activity, and downstream transcriptional programs. PROK2 contributes to neuroendocrine and circadian-related signaling, neuronal migration and survival, and modulation of inflammatory cell chemotaxis and vascular responses. In the central nervous system, prokineticin-2 is linked to olfactory bulb development and hypothalamic control of reproduction via GnRH neuron function, while in peripheral tissues it can shape leukocyte trafficking and pain-associated pathways. Altered PROK2/PROKR signaling has been associated with congenital hypogonadotropic hypogonadism and Kallmann syndrome, and is frequently examined in studies of neuroinflammation, angiogenic regulation, and tumor microenvironment signaling without implying clinical outcomes.

Prokineticin-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PROK2 expression without altering the underlying DNA sequence.

Prokineticin-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PROK2 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 PROK2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Prokineticin-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PROK2 locus and enabling the study of Prokineticin-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Prokineticin-2 pathway restoration in tumor cells with silenced or reduced PROK2 expression.

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