CHMP4C CRISPR Activation Plasmid (h): sc-404325-ACT

CHMP4C encodes a core component of the ESCRT-III machinery that remodels membranes during late stages of endosomal sorting, multivesicular body formation, and cytokinetic abscission. CHMP4C integrates into the abscission checkpoint to help coordinate final cell separation with chromosome segregation and genome integrity, linking ESCRT function to mitotic progression and DNA damage-associated signaling. Dysregulated ESCRT-III activity can perturb receptor downregulation, vesicle trafficking, and cell division fidelity, processes frequently studied in the context of tumorigenesis and chromosomal instability. As a result, CHMP4C is commonly investigated in pathways governing membrane scission, cell cycle control, and stress responses that influence proliferative and genome maintenance phenotypes.

CHMP4C CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CHMP4C expression without altering the underlying DNA sequence.

CHMP4C CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CHMP4C 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 CHMP4C transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CHMP4C expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CHMP4C locus and enabling the study of CHMP4C-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CHMP4C pathway restoration in tumor cells with silenced or reduced CHMP4C expression.

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