TRα CRISPR Activation Plasmid (h): sc-401475-ACT

THRA encodes thyroid hormone receptor alpha (TRα), a ligand-activated nuclear receptor that binds thyroid hormone response elements to regulate transcription programs controlling cellular metabolism, proliferation, differentiation, and development. TRα signaling integrates with coregulator-dependent chromatin remodeling and intersects with pathways governing mitochondrial function, cell cycle control, and tissue-specific gene expression. Dysregulated thyroid hormone receptor activity and altered THRA expression have been associated with developmental abnormalities and endocrine-related phenotypes, and TRα-dependent transcriptional programs are frequently studied in contexts such as cardiac, skeletal, and neural biology. As a transcription factor node, THRA is also used to interrogate hormone-responsive gene networks and epigenetic mechanisms of gene regulation in human cells.

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

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

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