Calpain 10 CRISPR Activation Plasmid (h): sc-405032-ACT

CAPN10 encodes calpain 10, a ubiquitously expressed, calcium-dependent cysteine protease that modulates limited proteolysis of intracellular substrates to regulate protein turnover, cytoskeletal remodeling, and signal transduction. Calpain 10 activity influences mitochondrial homeostasis, glucose and lipid metabolism, and stress-responsive pathways through proteolytic control of key metabolic and signaling proteins. Genetic variation and altered CAPN10 expression have been associated with metabolic phenotypes, including insulin resistance and type 2 diabetes susceptibility, supporting its relevance to studies of endocrine and mitochondrial dysfunction. In biomedical research, CAPN10 is frequently investigated for its roles in cellular energetics, inflammation-linked metabolic remodeling, and tissue-specific regulation of protease networks.

Calpain 10 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CAPN10 expression without altering the underlying DNA sequence.

Calpain 10 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CAPN10 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 CAPN10 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Calpain 10 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CAPN10 locus and enabling the study of Calpain 10-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Calpain 10 pathway restoration in tumor cells with silenced or reduced CAPN10 expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.