PRL-R CRISPR Activation Plasmid (h): sc-400814-ACT

PRLR encodes the human prolactin receptor (PRL-R), a type I cytokine receptor that mediates prolactin signaling to regulate epithelial differentiation, lactogenic gene expression, and tissue homeostasis. Ligand-dependent receptor dimerization activates JAK2/STAT5 as well as PI3K–AKT and MAPK/ERK pathways, integrating transcriptional and survival programs across endocrine-responsive cell types. Dysregulated PRL-R signaling has been linked to altered proliferation, differentiation, and inflammatory crosstalk in multiple tissues, making PRLR a widely studied node in hormone-driven biology. PRLR expression and pathway output are frequently interrogated in models of breast and prostate biology, metabolic regulation, and immune modulation to clarify context-specific signaling dependencies.

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

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

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