SMRTe CRISPR Activation Plasmid (h): sc-401150-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- SMRTe CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- SMRTe 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 SMRTe CRISPR Activation Plasmid (h) and SMRTe CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the NCOR2 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: SMRTe Antibody (1542/H7): sc-13554
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
SMRTe CRISPR Activation Plasmid (h) | sc-401150-ACT | 20 µg | $397.00 |
NCOR2 encodes SMRTe (silencing mediator for retinoid and thyroid hormone receptors), a corepressor that scaffolds transcriptional repression complexes containing HDAC3 and related chromatin-modifying factors. By engaging nuclear receptors and additional transcription factors, SMRTe helps coordinate chromatin compaction and context-dependent gene silencing across hormone-responsive and developmental gene programs. This regulation intersects with pathways controlling cell fate decisions, metabolic homeostasis, and inflammatory signaling through broad effects on enhancer and promoter acetylation. Dysregulated NCOR2/SMRTe activity has been implicated in altered transcriptional networks observed in cancer biology, endocrine-related phenotypes, and immune-mediated disease mechanisms, supporting its use as a mechanistic node in gene regulation studies.
SMRTe CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NCOR2 expression without altering the underlying DNA sequence.
SMRTe CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NCOR2 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 NCOR2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SMRTe expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NCOR2 locus and enabling the study of SMRTe-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SMRTe pathway restoration in tumor cells with silenced or reduced NCOR2 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.