TR2 CRISPR Activation Plasmid (h): sc-403818-ACT

Human NR2C1 encodes TR2, an orphan nuclear receptor that functions as a sequence-specific transcription factor regulating gene expression programs linked to cell fate, metabolism, and endocrine signaling. TR2 binds nuclear receptor response elements and interfaces with coregulators to modulate transcriptional networks involved in differentiation, reproductive biology, and developmental processes. In cellular pathways, TR2 activity can influence chromatin state and transcriptional homeostasis, shaping lineage decisions and stress-responsive gene expression. Dysregulated NR2C1/TR2 expression or signaling has been associated in the literature with altered differentiation states and cancer-relevant transcriptional programs, supporting its use as a mechanistic node in gene regulation studies.

TR2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NR2C1 expression without altering the underlying DNA sequence.

TR2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NR2C1 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 NR2C1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TR2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NR2C1 locus and enabling the study of TR2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TR2 pathway restoration in tumor cells with silenced or reduced NR2C1 expression.

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