transgelin CRISPR Activation Plasmid (h): sc-401343-ACT

TAGLN encodes transgelin, an actin-binding protein enriched in smooth muscle cells that stabilizes and bundles filamentous actin to support cytoskeletal architecture, contractility, and cell shape. Transgelin participates in actin cytoskeleton remodeling downstream of RhoA/ROCK and related mechanotransduction programs that coordinate adhesion and migration. Changes in TAGLN expression are frequently used as a marker of smooth muscle differentiation and phenotypic switching in vascular remodeling and fibrosis-associated processes. Dysregulated transgelin levels have also been reported in tumor-associated stromal biology and invasive cell states, linking cytoskeletal reprogramming to disease-relevant cellular behaviors.

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

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

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