FTβ CRISPR Activation Plasmid (h): sc-403479-ACT

FNTB encodes the beta subunit of protein farnesyltransferase, a key enzyme in the mevalonate/isoprenoid biosynthesis pathway that catalyzes farnesylation of CaaX motif–containing proteins. This lipid modification promotes membrane association and regulates localization and signaling of substrates such as RAS family GTPases and nuclear lamins, influencing proliferation, vesicular trafficking, and cytoskeletal dynamics. FTβ functions with the alpha subunit to form the active heterodimer and supports multiple small GTPase–driven pathways including MAPK and PI3K signaling outputs. Dysregulated prenylation and altered farnesyltransferase activity have been linked to oncogenic signaling networks and lamin-associated nuclear defects, making FNTB a relevant node for mechanistic studies in cancer biology and cell stress responses.

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

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

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