FAT2 CRISPR Activation Plasmid (h): sc-402931-ACT

Human FAT2 encodes an atypical cadherin-like protein in the FAT family implicated in cell–cell adhesion, planar cell polarity, and regulation of cytoskeletal organization. FAT2-associated signaling interfaces with processes that shape epithelial architecture, directional migration, and tissue morphogenesis, linking it to pathways governing polarity establishment and junctional integrity. Altered FAT family activity has been associated with dysregulated growth control and invasive phenotypes in multiple disease contexts, supporting investigation of FAT2 in mechanobiology and epithelial homeostasis. FAT2 is therefore relevant for studies of polarity-dependent signaling, membrane organization, and adhesion-linked transcriptional programs.

FAT2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FAT2 expression without altering the underlying DNA sequence.

FAT2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FAT2 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 FAT2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FAT2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FAT2 locus and enabling the study of FAT2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FAT2 pathway restoration in tumor cells with silenced or reduced FAT2 expression.

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