GR/NR3C1/Glucocorticoid Receptor CRISPR Activation Plasmid (h): sc-400071-ACT

NR3C1 encodes the glucocorticoid receptor (GR), a ligand-activated nuclear receptor that translocates to the nucleus upon hormone binding and regulates transcription through glucocorticoid response elements as well as tethering interactions with factors such as NF-κB and AP-1. GR signaling integrates endocrine cues with chromatin remodeling to control inflammatory gene programs, metabolic homeostasis, cellular differentiation, and stress responses. Through crosstalk with MAPK, PI3K–AKT, and cytokine signaling pathways, NR3C1 helps tune cell-type-specific transcriptional networks and feedback regulation of the hypothalamic–pituitary–adrenal axis. Dysregulated NR3C1 expression or function has been linked to altered glucocorticoid responsiveness and is frequently studied in contexts of immune dysregulation, neuropsychiatric stress biology, and cancer cell adaptation to microenvironmental stressors.

GR/NR3C1/Glucocorticoid Receptor CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NR3C1 expression without altering the underlying DNA sequence.

GR/NR3C1/Glucocorticoid Receptor CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NR3C1 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 NR3C1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GR/NR3C1/Glucocorticoid Receptor expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NR3C1 locus and enabling the study of GR/NR3C1/Glucocorticoid Receptor-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GR/NR3C1/Glucocorticoid Receptor pathway restoration in tumor cells with silenced or reduced NR3C1 expression.

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