Filamin 1 CRISPR Activation Plasmid (h): sc-400557-ACT

FLNA encodes filamin A, a large actin-binding scaffold that crosslinks F-actin and coordinates the assembly of mechanosensitive signaling complexes at the plasma membrane and along the cytoskeleton. Filamin A integrates cues from integrins, Rho family GTPases, and receptor-associated pathways to regulate cell shape, migration, adhesion dynamics, and membrane trafficking. Through interactions with channels, GPCRs, and signaling adaptors, it influences cytoskeletal remodeling and transcriptional responses to mechanical stress. Dysregulation or mutation of FLNA is linked to developmental disorders and contributes to altered cell motility and tissue architecture relevant to neurovascular and connective tissue phenotypes.

Filamin 1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FLNA expression without altering the underlying DNA sequence.

Filamin 1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FLNA 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 FLNA transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Filamin 1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FLNA locus and enabling the study of Filamin 1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Filamin 1 pathway restoration in tumor cells with silenced or reduced FLNA expression.

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