AKAP 12 CRISPR Activation Plasmid (h): sc-406008-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- AKAP 12 CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- AKAP 12 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 AKAP 12 CRISPR Activation Plasmid (h) and AKAP 12 CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the AKAP12 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: AKAP 12 Antibody (C-12): sc-376740
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
AKAP 12 CRISPR Activation Plasmid (h) | sc-406008-ACT | 20 µg | $397.00 | |||
AKAP 12 CRISPR Activation Plasmid (h2) | sc-406008-ACT-2 | 20 µg | $397.00 |
AKAP12 (A-kinase anchoring protein 12; gravin) is a scaffolding protein that spatially organizes PKA and PKC signaling at the plasma membrane and cytoskeleton, coordinating receptor desensitization, cytoskeletal remodeling, and cell-cycle control. By anchoring kinases and phosphatases near GPCRs and integrins, AKAP12 influences pathways governing focal adhesion dynamics, cell migration, and barrier function, with downstream effects on MAPK and cAMP-dependent signaling. Altered AKAP12 expression and epigenetic regulation have been linked to changes in cellular adhesion and motility programs relevant to cancer biology, vascular dysfunction, and inflammatory microenvironments. These properties make AKAP12 a useful node for studying signal compartmentalization and context-dependent transcriptional responses in human cells.
AKAP 12 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous AKAP12 expression without altering the underlying DNA sequence.
AKAP 12 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the AKAP12 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 AKAP12 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous AKAP 12 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native AKAP12 locus and enabling the study of AKAP 12-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of AKAP 12 pathway restoration in tumor cells with silenced or reduced AKAP12 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.