FAF1 CRISPR Activation Plasmid (h): sc-405535-ACT

FAF1 (Fas-associated factor 1) encodes a cytoplasmic adaptor protein that couples death receptor signaling to downstream apoptotic and proteostatic pathways. FAF1 interacts with FAS and caspase regulators and also modulates NF-κB signaling, influencing cell survival, inflammatory transcriptional programs, and stress responses. Through reported associations with ubiquitin–proteasome system components, FAF1 contributes to protein quality control and can shape cellular sensitivity to extrinsic apoptosis. Dysregulated FAF1 expression or function has been linked in the literature to cancer biology, neurodegenerative processes, and immune-related phenotypes, making it a useful node for mechanistic studies of cell fate decisions.

FAF1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FAF1 expression without altering the underlying DNA sequence.

FAF1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FAF1 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 FAF1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FAF1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FAF1 locus and enabling the study of FAF1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FAF1 pathway restoration in tumor cells with silenced or reduced FAF1 expression.

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