Androgen Receptor CRISPR Activation Plasmid (h): sc-400026-ACT

AR encodes the human androgen receptor, a ligand-activated nuclear hormone receptor that binds androgens, translocates to the nucleus, and regulates transcription through androgen response elements. Androgen receptor signaling coordinates cell-cycle progression, differentiation programs, metabolism, and chromatin remodeling via interactions with co-regulators and crosstalk with PI3K/AKT, MAPK, and WNT pathways. Dysregulated AR activity is linked to endocrine-driven biology and is frequently studied in hormone-responsive cancers, neuromuscular phenotypes such as spinal and bulbar muscular atrophy, and steroid metabolism disorders. As a transcriptional hub, AR provides a tractable readout for studying enhancer usage, epigenetic state, and stimulus-dependent gene networks.

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

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

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