FAM198B CRISPR Activation Plasmid (h): sc-409269-ACT

FAM198B (family with sequence similarity 198 member B) encodes a poorly characterized human protein with emerging links to membrane-associated processes and intracellular signaling that influence cell state and tissue organization. Transcriptomic studies have reported context-dependent regulation of FAM198B across multiple cell types, suggesting potential roles in pathways governing differentiation, adhesion, and stress-responsive transcriptional programs. Altered FAM198B expression has been observed in disease-associated datasets, supporting its utility as a molecular node for studying gene regulation in pathophysiology. Because mechanistic annotations remain limited, controlled modulation of endogenous FAM198B provides a tractable approach to map downstream gene networks and phenotypes.

FAM198B CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FAM198B expression without altering the underlying DNA sequence.

FAM198B CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FAM198B 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 FAM198B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FAM198B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FAM198B locus and enabling the study of FAM198B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FAM198B pathway restoration in tumor cells with silenced or reduced FAM198B expression.

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