EHD2 CRISPR Activation Plasmid (h): sc-403776-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- EHD2 CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- EHD2 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 EHD2 CRISPR Activation Plasmid (h) and EHD2 CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the EHD2 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: EHD2 Antibody (G-3): sc-515458
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
EHD2 CRISPR Activation Plasmid (h) | sc-403776-ACT | 20 µg | $397.00 |
EHD2 (EH domain containing 2) is a dynamin-related ATPase that associates with caveolae and the cortical actin cytoskeleton to regulate membrane curvature, endocytic trafficking, and caveolar stability. By constraining caveolae dynamics and influencing vesicle scission and recycling, EHD2 contributes to mechanotransduction and lipid handling pathways that shape cell adhesion, migration, and membrane tension responses. Dysregulated caveolar trafficking and cytoskeletal coupling involving EHD2 has been linked in the literature to altered metabolic homeostasis and oncogenic phenotypes, including changes in cellular invasiveness and signaling compartmentalization. As a result, EHD2 is frequently investigated in studies of endocytosis, plasma membrane organization, and caveolae-dependent signaling in human cells.
EHD2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EHD2 expression without altering the underlying DNA sequence.
EHD2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EHD2 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 EHD2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous EHD2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EHD2 locus and enabling the study of EHD2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of EHD2 pathway restoration in tumor cells with silenced or reduced EHD2 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.