copine 8 CRISPR Activation Plasmid (m): sc-426239-ACT

Mouse Cpne8 encodes copine 8, a calcium-dependent phospholipid-binding protein containing C2 domains and an A domain that supports regulated membrane association. Copine family members are implicated in calcium-triggered signaling events at the plasma membrane and endomembrane compartments, influencing vesicular trafficking, membrane remodeling, and protein recruitment dynamics. Through coupling Ca2+ flux to membrane-proximal protein interactions, copine 8 is relevant to pathways that shape receptor turnover and intracellular signaling amplitude. Altered regulation of calcium and membrane trafficking networks is frequently studied in contexts such as neuronal communication, immune signaling, and stress responses, making Cpne8 a useful target for mechanistic pathway interrogation.

copine 8 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Cpne8 expression without altering the underlying DNA sequence.

copine 8 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Cpne8 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 Cpne8 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous copine 8 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Cpne8 locus and enabling the study of copine 8-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of copine 8 pathway restoration in tumor cells with silenced or reduced Cpne8 expression.

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