folliculin CRISPR Activation Plasmid (h): sc-403316-ACT

Human FLCN encodes folliculin, a tumor suppressor–associated protein that functions as a GTPase-activating protein for RagC/D and helps couple amino acid sensing to mTORC1 signaling on lysosomes. Folliculin participates in energy and nutrient homeostasis through interactions with FNIP1/2 and AMPK, influencing autophagy, lysosomal biogenesis, and mitochondrial metabolism. Disruption of FLCN perturbs epithelial and stromal cell programs linked to proliferation and differentiation, and germline variants are associated with Birt–Hogg–Dubé syndrome, characterized by renal and pulmonary pathology. Because folliculin integrates metabolic and stress-response signaling, it is widely studied in pathways governing cellular growth control and tissue homeostasis.

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

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

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