FAM13A CRISPR Activation Plasmid (h): sc-405714-ACT

FAM13A (family with sequence similarity 13 member A) encodes a cytoplasmic protein implicated in regulation of Rho family GTPase signaling and cytoskeletal dynamics, with reported roles in epithelial cell polarity, migration, and differentiation. Genetic and functional studies link FAM13A to pathways shaping airway and alveolar homeostasis, including cell adhesion and remodeling processes that influence barrier function. Variation in FAM13A has been associated with lung function traits and chronic respiratory disease susceptibility, supporting its use as a molecular entry point for studying airway biology. In cancer and fibrosis-related contexts, altered FAM13A expression has been examined for effects on proliferation and motility programs, making it relevant for mechanistic studies of tissue remodeling.

FAM13A CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FAM13A expression without altering the underlying DNA sequence.

FAM13A CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FAM13A 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 FAM13A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FAM13A expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FAM13A locus and enabling the study of FAM13A-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FAM13A pathway restoration in tumor cells with silenced or reduced FAM13A expression.

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