Elongin B CRISPR Activation Plasmid (h): sc-402154-ACT

ELOB encodes human Elongin B, a core component of the Elongin BC complex that stabilizes SOCS-box and VHL-like adaptor proteins and supports assembly of Cullin-RING E3 ubiquitin ligases. Through these complexes, Elongin B contributes to ubiquitin-dependent proteostasis and regulation of transcriptional programs linked to RNA polymerase II elongation and oxygen-sensing pathways. ELOB-associated ligase assemblies influence turnover of key signaling and cell-cycle regulators, connecting Elongin B function to cellular stress responses, proliferation control, and genome integrity. Dysregulation of Elongin BC–dependent ubiquitination has been implicated in oncogenic signaling and altered hypoxia responses, making ELOB a useful node for mechanistic studies of protein degradation networks.

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

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

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