CHMP4A CRISPR Activation Plasmid (h): sc-402539-ACT

Human CHMP4A encodes a core component of the ESCRT-III machinery that remodels membranes to drive intraluminal vesicle formation during multivesicular body biogenesis and endosomal cargo sorting. CHMP4A also participates in cytokinetic abscission and helps coordinate membrane scission events important for plasma membrane repair and receptor downregulation. Through these roles, CHMP4A influences proteostasis and signaling pathway attenuation, including trafficking-dependent control of growth factor receptor outputs. Dysregulated ESCRT function is broadly linked to oncogenic signaling, neurodegenerative processes, and susceptibility to pathogens that exploit ESCRT-dependent budding, making CHMP4A a useful node for mechanistic studies of membrane remodeling.

CHMP4A CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CHMP4A expression without altering the underlying DNA sequence.

CHMP4A CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CHMP4A 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 CHMP4A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CHMP4A expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CHMP4A locus and enabling the study of CHMP4A-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CHMP4A pathway restoration in tumor cells with silenced or reduced CHMP4A expression.

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