Calpain reg CRISPR Activation Plasmid (h): sc-403478-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Calpain reg CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- Calpain reg 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 Calpain reg CRISPR Activation Plasmid (h) and Calpain reg CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the CAPNS1 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: Calpain reg Antibody (P-1): sc-32325
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Calpain reg CRISPR Activation Plasmid (h) | sc-403478-ACT | 20 µg | $397.00 |
CAPNS1 encodes the small regulatory subunit essential for stability and activity of the ubiquitous calpain-1 and calpain-2 proteases, enabling calcium-dependent, limited proteolysis of signaling and structural proteins. Through controlled substrate cleavage, CAPNS1-supported calpain activity modulates cytoskeletal remodeling, focal adhesion turnover, cell migration, and cell-cycle progression, and intersects with proteostasis and stress-response programs. Calpain signaling also influences inflammatory and apoptotic pathways by reshaping key regulators, linking CAPNS1 function to processes relevant to tumor biology, neurodegeneration, and tissue injury models. Altered CAPNS1 expression or calpain dysregulation has been associated with changes in cellular invasiveness, barrier integrity, and susceptibility to proteotoxic stress, making it a useful node for mechanistic studies.
Calpain reg CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CAPNS1 expression without altering the underlying DNA sequence.
Calpain reg CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CAPNS1 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 CAPNS1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Calpain reg expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CAPNS1 locus and enabling the study of Calpain reg-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Calpain reg pathway restoration in tumor cells with silenced or reduced CAPNS1 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.