FAT3 CRISPR Activation Plasmid (h): sc-404132-ACT

FAT3 encodes a large atypical cadherin that contributes to calcium-dependent cell–cell adhesion and helps organize tissue architecture through regulation of cell polarity, migration, and cytoskeletal dynamics. As a member of the FAT cadherin family, FAT3 is implicated in contact-dependent signaling programs that intersect with planar cell polarity and adhesion-associated pathways controlling morphogenesis and neuronal patterning. Altered FAT3 expression or regulatory disruption has been associated with changes in cellular motility and differentiation states, making it relevant for studying adhesion-driven remodeling in development and disease contexts. In human model systems, FAT3 serves as a useful node for investigating how membrane-anchored adhesion receptors influence transcriptional outputs and cell behavior.

FAT3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FAT3 expression without altering the underlying DNA sequence.

FAT3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FAT3 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 FAT3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FAT3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FAT3 locus and enabling the study of FAT3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FAT3 pathway restoration in tumor cells with silenced or reduced FAT3 expression.

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