TRB-1 CRISPR Activation Plasmid (h): sc-405781-ACT

Human TRIB1 (TRB-1) is a catalytically inactive pseudokinase that functions as a scaffold regulating signal transduction and protein turnover, notably through interactions with MAPK modules and ubiquitin ligases such as COP1. By modulating phosphorylation-dependent signaling outputs and stability of key transcriptional regulators, TRIB1 influences cell proliferation, differentiation, and stress-responsive programs. TRIB1-dependent networks are implicated in immune and inflammatory regulation and lipid metabolic homeostasis, and dysregulated expression has been associated with hematologic malignancy biology and cardiometabolic disease traits. These attributes make TRIB1 a useful target for dissecting pathway wiring and transcriptional state control in human cell models.

TRB-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TRIB1 expression without altering the underlying DNA sequence.

TRB-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TRIB1 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 TRIB1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TRB-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TRIB1 locus and enabling the study of TRB-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TRB-1 pathway restoration in tumor cells with silenced or reduced TRIB1 expression.

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