Talin CRISPR Activation Plasmid (h): sc-400783-ACT

Human TLN2 encodes talin-2, a large cytoskeletal adaptor that links integrin β cytoplasmic tails to F-actin, promoting focal adhesion assembly and transducing mechanical cues into intracellular signals. Through regulation of integrin activation, traction force generation, and adhesion turnover, talin-2 influences cell migration, tissue architecture, and mechanosensitive pathways that intersect with FAK/Src and Rho GTPase signaling. TLN2 activity is particularly relevant in contexts requiring robust adhesion and force transmission, including muscle and cardiac biology, and its dysregulation has been associated with altered cell motility and invasive phenotypes in cancer model systems. These properties make TLN2 a useful target for studying adhesion-dependent signaling, cytoskeletal remodeling, and mechanotransduction-driven transcriptional programs.

Talin-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TLN2 expression without altering the underlying DNA sequence.

Talin-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TLN2 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 TLN2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Talin-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TLN2 locus and enabling the study of Talin-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Talin-2 pathway restoration in tumor cells with silenced or reduced TLN2 expression.

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