KCC4 CRISPR Activation Plasmid (m): sc-422974-ACT
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- KCC4 CRISPR Activation Plasmid (m) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- KCC4 CRISPR Activation Plasmid (m) 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 KCC4 CRISPR Activation Plasmid (m) and KCC4 CRISPR Activation Plasmid (m2) target distinct regulatory regions upstream of the Slc12a7 transcriptional start site. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
KCC4 CRISPR Activation Plasmid (m) | sc-422974-ACT | 20 µg | $397.00 |
Slc12a7 encodes the mouse KCC4 (SLC12A7) K+-Cl− cotransporter, a key regulator of electroneutral chloride extrusion that helps set intracellular Cl− levels, cell volume, and ionic homeostasis. By shaping Cl− gradients, KCC4 influences membrane excitability and inhibitory signaling and intersects with osmotic stress responses and cytoskeletal remodeling programs linked to cell migration and invasion. KCC4 activity is also coupled to epithelial ion transport and pH/volume regulation, processes that can affect tissue barrier function and microenvironmental signaling. Dysregulated SLC12A7 expression has been associated with altered motility and remodeling phenotypes in disease-relevant contexts, supporting its use in mechanistic studies of ion transport–dependent cellular behaviors.
KCC4 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Slc12a7 expression without altering the underlying DNA sequence.
KCC4 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Slc12a7 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 Slc12a7 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous KCC4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Slc12a7 locus and enabling the study of KCC4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of KCC4 pathway restoration in tumor cells with silenced or reduced Slc12a7 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.