Talin CRISPR Activation Plasmid (h): sc-401342-ACT

Human TLN1 encodes talin, a cytoskeletal adaptor that links β-integrin cytoplasmic tails to F-actin to promote integrin activation, focal adhesion assembly, and mechanotransduction. Through coordination with focal adhesion kinase (FAK), Src-family signaling, Rho GTPase–dependent actin remodeling, and PI3K–AKT pathways, talin helps regulate cell adhesion, spreading, migration, and survival in response to extracellular matrix cues. Altered TLN1 expression or talin-dependent adhesion dynamics has been associated with dysregulated invasion and metastatic behavior, as well as defects in hemostasis and immune cell trafficking that depend on integrin signaling. These properties make TLN1 a widely used node for studying adhesion-dependent signaling networks and force-sensitive cell–matrix interactions.

Talin-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TLN1 expression without altering the underlying DNA sequence.

Talin-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TLN1 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 TLN1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Talin-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TLN1 locus and enabling the study of Talin-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Talin-1 pathway restoration in tumor cells with silenced or reduced TLN1 expression.

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