CCDC89 CRISPR Activation Plasmid (h): sc-408069-ACT

CCDC89 (coiled-coil domain containing 89) encodes a predicted coiled-coil protein implicated in cytoskeletal organization and protein–protein interaction networks that support intracellular architecture and signaling. Coiled-coil proteins frequently participate in scaffolding functions that influence organelle positioning, vesicular trafficking, and stress-responsive pathways, suggesting CCDC89 may modulate spatial coordination of cellular processes. Expression- and variant-level associations reported across genomics datasets indicate potential relevance of CCDC89 to dysregulated transcriptional programs observed in proliferative and neurobiological contexts, motivating mechanistic studies in disease-relevant cell models. Defining how CCDC89 impacts pathway wiring can clarify downstream effects on cell state transitions, adhesion/motility phenotypes, and proteostasis.

CCDC89 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CCDC89 expression without altering the underlying DNA sequence.

CCDC89 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CCDC89 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 CCDC89 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CCDC89 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CCDC89 locus and enabling the study of CCDC89-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CCDC89 pathway restoration in tumor cells with silenced or reduced CCDC89 expression.

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