karyopherin α6 CRISPR Activation Plasmid (h): sc-403459-ACT

KPNA6 encodes karyopherin alpha 6, an importin-α family adaptor that recognizes classical nuclear localization signals and couples cargo proteins to importin-β for transport through the nuclear pore complex. By regulating nucleo-cytoplasmic trafficking of transcription factors, DNA repair proteins, and cell-cycle regulators, KPNA6 contributes to control of gene expression programs and proteostasis. Altered nuclear import dynamics are implicated in dysregulated proliferation, stress responses, and genome stability, linking KPNA6-dependent transport to mechanisms relevant in cancer biology and other disorders where nuclear signaling is perturbed. KPNA6 is therefore a useful node for studying compartmentalization-dependent regulation of transcription and signaling pathways.

karyopherin α6 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KPNA6 expression without altering the underlying DNA sequence.

karyopherin α6 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KPNA6 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 KPNA6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous karyopherin α6 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KPNA6 locus and enabling the study of karyopherin α6-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of karyopherin α6 pathway restoration in tumor cells with silenced or reduced KPNA6 expression.

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