RARα/Retinoic Acid Receptor α CRISPR Activation Plasmid (h): sc-400529-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- RARα/Retinoic Acid Receptor α CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- RARα/Retinoic Acid Receptor α 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 RARα/Retinoic Acid Receptor α CRISPR Activation Plasmid (h) and RARα/Retinoic Acid Receptor α CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the RARA 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: RARα/Retinoic Acid Receptor α Antibody (C-1): sc-515796
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
RARα/Retinoic Acid Receptor α CRISPR Activation Plasmid (h) | sc-400529-ACT | 20 µg | $397.00 | |||
RARα/Retinoic Acid Receptor α CRISPR Activation Plasmid (h2) | sc-400529-ACT-2 | 20 µg | $397.00 |
RARA encodes RARα, a ligand-activated nuclear receptor that heterodimerizes with RXRs to bind retinoic acid response elements and regulate transcription programs controlling cell fate decisions. Through retinoid signaling, RARα modulates chromatin state and gene networks governing differentiation, proliferation, and apoptosis, with strong links to hematopoietic maturation and epithelial development. Its activity integrates with core transcriptional and epigenetic pathways, including co-repressor/co-activator exchange and cross-talk with MAPK and PI3K signaling. Dysregulated RARA expression or signaling is associated with altered differentiation states and is widely studied in cancer biology and developmental disease models.
RARα/Retinoic Acid Receptor α CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RARA expression without altering the underlying DNA sequence.
RARα/Retinoic Acid Receptor α CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RARA 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 RARA transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous RARα/Retinoic Acid Receptor α expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RARA locus and enabling the study of RARα/Retinoic Acid Receptor α-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of RARα/Retinoic Acid Receptor α pathway restoration in tumor cells with silenced or reduced RARA expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.