Cofilin 1 CRISPR Activation Plasmid (h): sc-400394-ACT

CFL1 encodes cofilin 1, a central regulator of actin filament turnover that binds F-actin and promotes severing and depolymerization to support rapid cytoskeletal remodeling. Its activity is tightly controlled by phosphorylation-dependent signaling and upstream pathways such as Rho GTPase–LIMK–SSH, coordinating lamellipodia formation, cell polarity, cytokinesis, and endocytosis. By shaping actin dynamics, cofilin 1 influences cell migration, adhesion, and mechanotransduction, processes frequently perturbed in cancer invasion and metastasis as well as in cardiovascular and inflammatory contexts. Dysregulated cofilin signaling and actin remodeling have also been linked to neurodegenerative phenotypes, including aberrant actin–cofilin aggregates reported in models of synaptic dysfunction.

Cofilin 1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CFL1 expression without altering the underlying DNA sequence.

Cofilin 1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CFL1 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 CFL1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Cofilin 1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CFL1 locus and enabling the study of Cofilin 1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Cofilin 1 pathway restoration in tumor cells with silenced or reduced CFL1 expression.

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