karyopherin β1 CRISPR Activation Plasmid (h): sc-401230-ACT

KPNB1 encodes karyopherin β1 (importin β1), a central nuclear transport receptor that binds importin α–cargo complexes and mediates Ran GTPase–dependent translocation through the nuclear pore complex. By controlling nuclear import of transcription factors, cell cycle regulators, and DNA damage response proteins, KPNB1 helps coordinate transcriptional programs, mitotic progression, and stress signaling. Altered nucleocytoplasmic trafficking is a recurring feature of proliferative and neurodegenerative phenotypes, and dysregulated KPNB1 expression or activity has been associated with aberrant growth control and genome maintenance pathways in multiple disease contexts.

karyopherin β1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KPNB1 expression without altering the underlying DNA sequence.

karyopherin β1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KPNB1 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 KPNB1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous karyopherin β1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KPNB1 locus and enabling the study of karyopherin β1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of karyopherin β1 pathway restoration in tumor cells with silenced or reduced KPNB1 expression.

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