CCDC23 CRISPR Activation Plasmid (h): sc-415702-ACT

SVBP encodes CCDC23, a coiled-coil domain–containing protein implicated in centrosome-associated processes and microtubule organization, with reported roles in regulating the mitotic spindle and chromosome segregation. Through these functions, CCDC23 is positioned to influence cell-cycle progression and genome stability pathways that are frequently perturbed in proliferative disease contexts. Altered expression or dysregulation of centrosomal and spindle-associated factors is commonly linked to aneuploidy and cellular stress responses, making SVBP/CCDC23 relevant for mechanistic studies in cancer biology and related models. Its intracellular localization and predicted scaffolding features also support use as a handle for probing protein interaction networks that govern mitosis and cytoskeletal dynamics.

CCDC23 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SVBP expression without altering the underlying DNA sequence.

CCDC23 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SVBP 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 SVBP transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CCDC23 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SVBP locus and enabling the study of CCDC23-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CCDC23 pathway restoration in tumor cells with silenced or reduced SVBP expression.

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