ERRβ CRISPR Activation Plasmid (h): sc-402868-ACT

ESRRB encodes estrogen-related receptor beta (ERRβ), an orphan nuclear receptor transcription factor that regulates gene programs controlling cellular energy homeostasis, mitochondrial biogenesis, and oxidative metabolism. ERRβ binds specific DNA response elements and interfaces with co-regulators to modulate transcriptional networks linked to differentiation and cell-state maintenance. In human cells, ESRRB activity intersects with nuclear receptor signaling and metabolic pathways, influencing processes such as proliferation, stress adaptation, and lineage specification. Dysregulated ERRβ-associated transcriptional control has been investigated in contexts including developmental disorders and cancer-relevant metabolic remodeling.

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

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

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