Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
DDIT3/CHOP/GADD153 CRISPR Activation Plasmid (h): sc-400051-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- DDIT3/CHOP/GADD153 CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- DDIT3/CHOP/GADD153 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 DDIT3/CHOP/GADD153 CRISPR Activation Plasmid (h) and DDIT3/CHOP/GADD153 CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the DDIT3 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: DDIT3/GADD153/CHOP Antibody (B-3): sc-7351
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
DDIT3/CHOP/GADD153 CRISPR Activation Plasmid (h) | sc-400051-ACT | 20 µg | $397.00 | |||
DDIT3/CHOP/GADD153 CRISPR Activation Plasmid (h2) | sc-400051-ACT-2 | 20 µg | $397.00 |
DDIT3, also known as CHOP/GADD153, encodes a stress-inducible bZIP transcription factor that integrates signals from the unfolded protein response and the integrated stress response. It is strongly induced downstream of PERK–eIF2α–ATF4 during endoplasmic reticulum stress and modulates transcriptional programs controlling apoptosis, autophagy, redox balance, and cell-cycle regulation. DDIT3 influences adaptive versus terminal stress outcomes by altering expression of genes involved in protein folding, amino acid metabolism, and mitochondrial function. Dysregulated DDIT3 signaling has been linked to cancer cell survival under proteotoxic stress, metabolic and neurodegenerative stress phenotypes, and oncogenic fusion events such as FUS–DDIT3 in myxoid liposarcoma.
DDIT3/CHOP/GADD153 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DDIT3 expression without altering the underlying DNA sequence.
DDIT3/CHOP/GADD153 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DDIT3 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 DDIT3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DDIT3/CHOP/GADD153 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DDIT3 locus and enabling the study of DDIT3/CHOP/GADD153-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DDIT3/CHOP/GADD153 pathway restoration in tumor cells with silenced or reduced DDIT3 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.