PDZ-RhoGEF CRISPR Activation Plasmid (h): sc-403444-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PDZ-RhoGEF CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- PDZ-RhoGEF 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 PDZ-RhoGEF CRISPR Activation Plasmid (h) and PDZ-RhoGEF CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the ARHGEF11 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: PDZ-RhoGEF Antibody (D-9): sc-166740
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PDZ-RhoGEF CRISPR Activation Plasmid (h) | sc-403444-ACT | 20 µg | $397.00 | |||
PDZ-RhoGEF CRISPR Activation Plasmid (h2) | sc-403444-ACT-2 | 20 µg | $397.00 |
ARHGEF11 encodes PDZ-RhoGEF, a RhoA-specific guanine nucleotide exchange factor that couples activated Gα12/13 signaling to RhoA activation and downstream cytoskeletal remodeling. By promoting RhoA–ROCK pathway output, PDZ-RhoGEF regulates actin stress fiber formation, focal adhesion dynamics, cell polarity, and contractility, influencing processes such as cell migration and junctional organization. ARHGEF11-linked signaling intersects with GPCR networks and mechanotransduction pathways that shape tissue architecture and inflammatory responses. Dysregulation of Rho GTPase signaling involving PDZ-RhoGEF has been associated with altered motility and invasiveness in cancer biology and with vascular and metabolic phenotypes in genetic and functional studies.
PDZ-RhoGEF CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ARHGEF11 expression without altering the underlying DNA sequence.
PDZ-RhoGEF CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ARHGEF11 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 ARHGEF11 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PDZ-RhoGEF expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ARHGEF11 locus and enabling the study of PDZ-RhoGEF-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PDZ-RhoGEF pathway restoration in tumor cells with silenced or reduced ARHGEF11 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.