GCN2 CRISPR Activation Plasmid (h): sc-402313-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GCN2 CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- GCN2 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 GCN2 CRISPR Activation Plasmid (h) and GCN2 CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the EIF2AK4 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: GCN2 Antibody (F-7): sc-374609
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
GCN2 CRISPR Activation Plasmid (h) | sc-402313-ACT | 20 µg | $397.00 |
EIF2AK4 encodes GCN2, a serine/threonine kinase that senses amino acid limitation through binding of uncharged tRNAs and couples nutrient stress to translational control. Upon activation, GCN2 phosphorylates eIF2α to initiate the integrated stress response, reshaping global protein synthesis while promoting selective translation programs that support adaptation to metabolic and oxidative stress. This signaling intersects with autophagy, mitochondrial homeostasis, and immune-related transcriptional outputs via ATF4-dependent pathways. Altered EIF2AK4/GCN2 activity has been studied in contexts of nutrient stress adaptation, inflammation, and tumor cell survival, making it relevant for dissecting stress-signaling dependencies in human cellular models.
GCN2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EIF2AK4 expression without altering the underlying DNA sequence.
GCN2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EIF2AK4 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 EIF2AK4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GCN2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EIF2AK4 locus and enabling the study of GCN2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GCN2 pathway restoration in tumor cells with silenced or reduced EIF2AK4 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.