karyopherin β3 CRISPR Activation Plasmid (h): sc-402071-ACT

IPO5 encodes the nuclear import receptor karyopherin β3, a RanGTP-dependent transportin family member that mediates selective translocation of protein cargos through the nuclear pore complex. By controlling nucleocytoplasmic trafficking, IPO5 influences transcriptional programs, cell-cycle progression, and stress-responsive signaling through regulation of nuclear availability of key regulatory proteins. Altered nuclear transport dynamics are frequently linked to oncogenic transformation, viral infection biology, and neurodegeneration, making IPO5 a relevant node for studying how cargo localization reshapes cellular phenotypes. In human cells, IPO5-dependent import can modulate pathway output by changing the nuclear residency of transcription factors and other signaling effectors.

karyopherin β3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous IPO5 expression without altering the underlying DNA sequence.

karyopherin β3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the IPO5 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 IPO5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous karyopherin β3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native IPO5 locus and enabling the study of karyopherin β3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of karyopherin β3 pathway restoration in tumor cells with silenced or reduced IPO5 expression.

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