CHTOP CRISPR Activation Plasmid (h): sc-411874-ACT

CHTOP (chromatin target of PRMT1) encodes a nuclear RNA-binding protein that functions as a cofactor in mRNA biogenesis and export, integrating chromatin-associated events with post-transcriptional gene regulation. It participates in the TREX mRNA export pathway and couples transcriptional output to efficient nuclear export through interactions with RNA processing and transport machinery. CHTOP is also linked to arginine methylation–dependent regulation and influences alternative splicing, ribonucleoprotein assembly, and genome-wide expression programs. Dysregulated CHTOP activity has been associated with altered RNA metabolism and proliferative signaling states relevant to cancer biology and other diseases characterized by disrupted gene expression control.

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

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

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