TGase2 CRISPR Activation Plasmid (h): sc-401543-ACT

Human TGM2 encodes transglutaminase 2 (TGase2), a multifunctional Ca2+-dependent enzyme that catalyzes protein crosslinking and polyamination and can also act as a GTP-binding signaling protein. TGase2 regulates cytoskeletal remodeling, cell adhesion, extracellular matrix stabilization, autophagy, and apoptosis, interfacing with integrin/FAK signaling, NF-κB–linked inflammatory programs, and TGF-β–associated remodeling pathways. Through these processes, altered TGase2 activity has been associated with fibrosis and aberrant matrix deposition, inflammatory states, and tumor cell survival and invasion phenotypes. Its broad subcellular localization and context-dependent functions make TGM2 a useful node for mechanistic studies of stress responses and cell fate control.

TGase2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TGM2 expression without altering the underlying DNA sequence.

TGase2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TGM2 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 TGM2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TGase2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TGM2 locus and enabling the study of TGase2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TGase2 pathway restoration in tumor cells with silenced or reduced TGM2 expression.

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