Filamin 2 CRISPR Activation Plasmid (h): sc-402358-ACT

Human FLNC encodes filamin 2, an actin-binding cytoskeletal scaffold that crosslinks F-actin and couples the cortical cytoskeleton to membrane proteins, coordinating cell shape, adhesion, and mechanotransduction. Filamin 2 participates in focal adhesion and integrin-associated signaling, influencing cytoskeletal remodeling, migration, and force transmission in muscle and other mechanically stressed tissues. FLNC-dependent networks interface with pathways controlling sarcomere integrity and stress responses, linking cytoskeletal organization to cellular homeostasis. Genetic disruption or dysregulated expression of FLNC is associated with cardiomyopathy and skeletal myopathies, supporting its relevance for studies of muscle biology and disease mechanisms.

Filamin 2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FLNC expression without altering the underlying DNA sequence.

Filamin 2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FLNC 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 FLNC transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Filamin 2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FLNC locus and enabling the study of Filamin 2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Filamin 2 pathway restoration in tumor cells with silenced or reduced FLNC expression.

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