TRβ CRISPR Activation Plasmid (h): sc-401467-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- TRβ CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- TRβ CRISPR Activation Plasmid (h) consists of three plasmids at a 1:1:1 mass ratio: a plasmid encoding the deactivated Cas9 (dCas9) nuclease (D10A and N863A) fused to the transactivation domain VP64, and a blasticidin resistance gene; a plasmid encoding the MS2-p65-HSF1 fusion protein, and a hygromycin resistance gene; a plasmid encoding a target-specific 20 nt guide RNA fused to two MS2 RNA aptamers, and a puromycin resistance gene
- The resulting SAM complex binds to a site-specific region approximately 200-250 nt upstream of the transcriptional start site and provides robust recruitment of transcription factors for highly efficient gene activation
- gRNAs encoded by TRβ CRISPR Activation Plasmid (h) and TRβ CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the THRB transcriptional start site. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: TRβ1 Antibody (J51): sc-737
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
TRβ CRISPR Activation Plasmid (h) | sc-401467-ACT | 20 µg | $397.00 | |||
TRβ CRISPR Activation Plasmid (h2) | sc-401467-ACT-2 | 20 µg | $397.00 |
THRB encodes thyroid hormone receptor beta (TRβ), a ligand-dependent nuclear receptor that binds triiodothyronine (T3) and regulates transcription through thyroid hormone response elements. TRβ coordinates gene programs controlling cellular metabolism, mitochondrial function, lipid and carbohydrate homeostasis, and developmental differentiation via interactions with coregulators and crosstalk with RXR and MAPK/PI3K signaling. Altered THRB activity is linked to dysregulated endocrine signaling and metabolic phenotypes, and perturbations in TRβ-mediated transcriptional networks are studied in contexts such as thyroid hormone resistance and hormone-responsive cancers. In human cells, TRβ serves as a model transcription factor for probing nuclear receptor chromatin occupancy, cofactor dependency, and stimulus-responsive gene regulation.
TRβ CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous THRB expression without altering the underlying DNA sequence.
TRβ CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the THRB 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 THRB transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TRβ expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native THRB locus and enabling the study of TRβ-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TRβ pathway restoration in tumor cells with silenced or reduced THRB expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.