β-TrCP CRISPR Activation Plasmid (h): sc-400971-ACT

BTRC encodes the human F-box protein β-TrCP, a substrate recognition component of the SCF (SKP1–CUL1–F-box) E3 ubiquitin ligase complex that directs phosphodegron-dependent ubiquitination and proteasomal turnover of diverse regulatory proteins. Through this activity, β-TrCP integrates signaling and proteostasis across pathways including NF-κB (via IκB regulation), Wnt/β-catenin, cell-cycle control, and DNA damage responses, thereby shaping transcriptional programs and checkpoint integrity. Altered BTRC expression or substrate targeting can perturb inflammatory signaling, proliferation, and genomic stability, processes frequently implicated in cancer biology and other complex disease mechanisms. Because β-TrCP has broad substrate scope, it is commonly studied to map ubiquitin-mediated network rewiring and pathway cross-talk under defined stimuli.

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

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

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