FTα CRISPR Activation Plasmid (h): sc-403920-ACT

FNTA encodes the alpha subunit of protein farnesyltransferase (FTα), which forms a heterodimeric prenyltransferase complex that catalyzes farnesylation of C-terminal CAAX motif proteins. This lipid modification supports membrane association, subcellular trafficking, and protein–protein interactions for key signaling regulators, including members of the RAS superfamily, thereby influencing pathways controlling proliferation, differentiation, and cytoskeletal organization. Altered prenylation dynamics and FNTA-dependent processing have been associated with dysregulated oncogenic signaling, aberrant cell growth programs, and defects in intracellular organization relevant to cancer biology and other proliferative disorders.

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

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

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