FAM186A CRISPR Activation Plasmid (h2): sc-414163-ACT-2

Human FAM186A encodes a poorly characterized protein implicated by expression and genetic association data in cell type–specific regulatory programs and tissue homeostasis. Although its precise molecular function remains unresolved, FAM186A is commonly studied in the context of intracellular signaling and transcriptional control networks that influence cellular differentiation and stress-adaptive responses. Variants or dysregulated expression of FAM186A have been reported in datasets linked to complex disease traits, motivating investigation of its contribution to pathogenic mechanisms through functional genomics. Gene editing of FAM186A supports loss- and gain-of-function studies, pathway mapping, and phenotypic profiling in human cell models to clarify downstream effects on cellular processes and disease-relevant molecular signatures.

FAM186A CRISPR Activation Plasmid (h2) provides a targeted, non-destructive approach to upregulating endogenous FAM186A expression without altering the underlying DNA sequence.

FAM186A CRISPR Activation Plasmid (h2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FAM186A 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 FAM186A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FAM186A expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FAM186A locus and enabling the study of FAM186A-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FAM186A pathway restoration in tumor cells with silenced or reduced FAM186A expression.

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