PTRF CRISPR Activation Plasmid (h): sc-404669-ACT

Human CAVIN1 (PTRF) encodes polymerase I and transcript release factor, a core cavin family component required for caveolae biogenesis and stabilization at the plasma membrane. PTRF coordinates caveolin-dependent membrane curvature, mechanoprotection, and caveolae-mediated endocytosis, influencing signal compartmentalization and lipid homeostasis. Through its role in organizing membrane microdomains, PTRF impacts pathways linked to insulin signaling, adipocyte differentiation, and cytoskeletal dynamics. Altered CAVIN1/PTRF function or expression has been associated with lipodystrophy-related phenotypes, muscular abnormalities, and dysregulated cell signaling relevant to cancer biology and metabolic disease research.

PTRF CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CAVIN1 expression without altering the underlying DNA sequence.

PTRF CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CAVIN1 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 CAVIN1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PTRF expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CAVIN1 locus and enabling the study of PTRF-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PTRF pathway restoration in tumor cells with silenced or reduced CAVIN1 expression.

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