RNF123 CRISPR Activation Plasmid (h): sc-403631-ACT

RNF123 (also known as KPC1) encodes a RING-type E3 ubiquitin ligase that helps control protein homeostasis by catalyzing ubiquitin-dependent turnover of specific substrates. It has been implicated in regulation of cell-cycle progression through ubiquitin-mediated processing of CDKN1B/p27 and related checkpoint circuitry, linking RNF123 activity to proliferation and stress-response programs. As part of the ubiquitin–proteasome system, RNF123 influences signaling outputs by adjusting the stability of regulatory proteins that govern growth and differentiation. Dysregulated ubiquitination pathways involving RNF123 are therefore of interest in studies of oncogenic signaling, genome stability, and proteostasis-associated disease mechanisms.

RNF123 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RNF123 expression without altering the underlying DNA sequence.

RNF123 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RNF123 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 RNF123 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous RNF123 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RNF123 locus and enabling the study of RNF123-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of RNF123 pathway restoration in tumor cells with silenced or reduced RNF123 expression.

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