QKI CRISPR Activation Plasmid (h): sc-405082-ACT

QKI (Quaking) is an RNA-binding protein that regulates post-transcriptional gene expression through control of pre-mRNA splicing, mRNA stabilization, export, and translation. It is a key modulator of cell fate programs, with prominent roles in oligodendrocyte differentiation and myelination, and it coordinates RNA regulons involved in cytoskeletal organization and signal transduction. QKI interfaces with pathways governing epithelial–mesenchymal dynamics, apoptosis, and stress responses by shaping transcript isoform usage and RNA turnover. Dysregulated QKI expression or splicing has been associated with neurodevelopmental and demyelinating phenotypes and is frequently studied in the context of cancer biology where altered RNA processing contributes to aberrant proliferation and invasion.

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

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

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