FRY CRISPR Activation Plasmid (h): sc-404629-ACT

Human FRY encodes a large, evolutionarily conserved microtubule-associated protein implicated in cytoskeletal organization, cell polarity, and control of proliferation. FRY has been linked to hippo-related signaling and kinase networks that couple centrosome and microtubule dynamics to cell-cycle progression and morphogenesis. Through these processes, FRY is studied in contexts such as epithelial organization, neuronal development, and migration, where perturbations in cytoskeletal regulation can influence oncogenic phenotypes. Altered FRY expression or function has been reported across multiple disease-relevant datasets, supporting its use as a research target for dissecting pathways that govern growth control and tissue architecture.

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

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

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