FNBP4 CRISPR Activation Plasmid (h): sc-408501-ACT

FNBP4 (formin binding protein 4) is a cytoplasmic adaptor implicated in coupling membrane dynamics to actin cytoskeleton remodeling through interactions with proline-rich and SH3-domain–containing partners. By linking signaling complexes to endocytic and cytoskeletal machinery, FNBP4 can influence vesicle trafficking, cell shape changes, and migration programs. Altered regulation of these processes is frequently associated with dysregulated growth and invasion phenotypes, making FNBP4 a relevant node for studying pathway rewiring in cancer and other disorders involving aberrant cytoskeletal control. Functional interrogation of FNBP4 supports mechanistic studies of actin-dependent signaling, membrane trafficking, and context-specific transcriptional outputs driven by these pathways.

FNBP4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FNBP4 expression without altering the underlying DNA sequence.

FNBP4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FNBP4 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 FNBP4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FNBP4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FNBP4 locus and enabling the study of FNBP4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FNBP4 pathway restoration in tumor cells with silenced or reduced FNBP4 expression.

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