β-FR CRISPR Activation Plasmid (h): sc-403346-ACT

FOLR2 encodes folate receptor beta (β-FR), a glycosylphosphatidylinositol-anchored cell-surface protein that binds reduced folates and mediates receptor-dependent uptake to support one-carbon metabolism. By regulating intracellular folate availability, β-FR influences nucleotide biosynthesis and methylation reactions that are tightly linked to cell proliferation and differentiation. FOLR2 is enriched in myeloid-lineage cells, including subsets of monocytes and macrophages, and is commonly used as a marker of macrophage state and tissue immune biology. Altered FOLR2 expression has been associated with inflammatory microenvironments and tumor-associated myeloid phenotypes, making it relevant to studies of immune regulation and cancer biology.

β-FR CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FOLR2 expression without altering the underlying DNA sequence.

β-FR CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FOLR2 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 FOLR2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous β-FR expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FOLR2 locus and enabling the study of β-FR-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of β-FR pathway restoration in tumor cells with silenced or reduced FOLR2 expression.

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