karyopherin α1 CRISPR Activation Plasmid (h): sc-402324-ACT

KPNA1 encodes karyopherin alpha 1, an importin-α adaptor that recognizes classical nuclear localization signals and couples cargo proteins to importin-β for Ran GTPase–regulated transport through the nuclear pore complex. By controlling nuclear entry of transcription factors, cell cycle regulators, and DNA damage response proteins, KPNA1 helps shape gene expression programs, chromatin regulation, and stress-adaptive signaling. Altered nucleocytoplasmic trafficking involving importin pathways has been linked to oncogenic signaling, impaired immune and inflammatory responses, and viral life cycle dependencies, making KPNA1 relevant for mechanistic studies of these processes. KPNA1 activity is also pertinent to proteostasis and nuclear compartmentalization phenotypes that influence differentiation and cellular identity.

karyopherin α1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KPNA1 expression without altering the underlying DNA sequence.

karyopherin α1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KPNA1 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 KPNA1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous karyopherin α1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KPNA1 locus and enabling the study of karyopherin α1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of karyopherin α1 pathway restoration in tumor cells with silenced or reduced KPNA1 expression.

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