RORβ CRISPR Activation Plasmid (h): sc-402834-ACT

RORB encodes the nuclear receptor RORβ, a DNA-binding transcription factor that regulates gene programs involved in neuronal differentiation, circadian rhythm output, and sensory system development. As a member of the ROR/REV-ERB network, RORβ interfaces with transcriptional control of metabolic and developmental pathways, shaping cell-type–specific identity and excitability. RORB expression is enriched in the central nervous system, including retinal and cortical populations, where it contributes to neural circuit formation and photoreceptor-related gene regulation. Dysregulation of RORB-associated transcriptional programs has been linked in research settings to neurodevelopmental and neuropsychiatric disease mechanisms.

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

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

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