QIP1 CRISPR Activation Plasmid (h): sc-405442-ACT

KPNA4 encodes karyopherin subunit alpha 4, also referred to as QIP1, a nuclear import adaptor that binds classical nuclear localization signals and partners with importin beta to traffic cargo proteins through the nuclear pore complex. By regulating nucleocytoplasmic transport, KPNA4 influences transcriptional programs, cell-cycle control, stress responses, and signaling pathways that depend on timely nuclear entry of transcription factors and regulatory enzymes. Altered importin-α family activity has been associated with dysregulated gene expression and aberrant growth signaling, making KPNA4/QIP1 relevant to studies of oncogenic signaling, viral–host interactions, and neurobiology where nuclear transport capacity can be rate-limiting. Mapping KPNA4-dependent cargo and transport dynamics is therefore useful for dissecting pathway connectivity and context-specific nuclear import requirements in human cells.

QIP1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KPNA4 expression without altering the underlying DNA sequence.

QIP1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KPNA4 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 KPNA4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous QIP1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KPNA4 locus and enabling the study of QIP1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of QIP1 pathway restoration in tumor cells with silenced or reduced KPNA4 expression.

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