CRBN CRISPR Activation Plasmid (h): sc-412142-ACT

Human CRBN encodes cereblon, a substrate receptor of the CUL4–DDB1 E3 ubiquitin ligase complex that controls protein homeostasis by directing specific targets for ubiquitination and proteasomal turnover. Through regulation of ubiquitin-dependent degradation, CRBN influences transcriptional programs, cell-cycle progression, and cellular stress responses, linking it to pathways that shape differentiation and immune signaling. CRBN has been studied in the context of developmental biology and neuronal function, and perturbation of its expression or activity is associated with altered proteostasis and disease-relevant phenotypes. As a central node in ubiquitin–proteasome system regulation, CRBN is frequently interrogated to understand context-specific substrate selection and downstream signaling consequences.

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

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

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