γ-FR CRISPR Activation Plasmid (h): sc-403347-ACT

Human FOLR3 encodes γ-folate receptor (γ-FR), a glycosylphosphatidylinositol-associated folate receptor family member implicated in folate binding and cellular folate homeostasis, supporting one-carbon metabolism required for nucleotide biosynthesis and methylation reactions. Through its connection to folate availability, γ-FR links to pathways influencing DNA replication and repair, redox balance, and epigenetic regulation, processes that are frequently perturbed in rapidly proliferating or metabolically reprogrammed cells. Variation in folate receptor expression has been explored in contexts of hematopoietic and immune cell biology and in diseases where folate metabolism and methylation status contribute to altered transcriptional programs. As a surface-associated folate-binding protein, γ-FR also provides a tractable handle for studying receptor-mediated nutrient handling and downstream metabolic phenotypes in human cell models.

γ-FR CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FOLR3 expression without altering the underlying DNA sequence.

γ-FR CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FOLR3 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 FOLR3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous γ-FR expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FOLR3 locus and enabling the study of γ-FR-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of γ-FR pathway restoration in tumor cells with silenced or reduced FOLR3 expression.

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