TRAP-1 CRISPR Activation Plasmid (h): sc-402963-ACT

Human TGFBRAP1 encodes TRAP-1, an accessory factor implicated in transforming growth factor-β (TGF-β) receptor signaling and intracellular trafficking processes that influence receptor maturation, stability, and downstream pathway propagation. By modulating the efficiency and context of SMAD-dependent transcriptional programs, TRAP-1 can affect cellular outcomes such as proliferation control, differentiation, extracellular matrix remodeling, and immune-related signaling cross-talk. Perturbations in TGF-β pathway dynamics are widely connected to fibrosis, cancer-associated signaling rewiring, and inflammatory disease biology, making TRAP-1 a useful node for mechanistic studies of pathway tuning. Expression and functional interrogation of TGFBRAP1 supports research into signal transduction architecture, receptor-associated complexes, and context-dependent transcriptional responses.

TRAP-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TGFBRAP1 expression without altering the underlying DNA sequence.

TRAP-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TGFBRAP1 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 TGFBRAP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TRAP-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TGFBRAP1 locus and enabling the study of TRAP-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TRAP-1 pathway restoration in tumor cells with silenced or reduced TGFBRAP1 expression.

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