Importin-7 CRISPR Activation Plasmid (h): sc-403562-ACT

IPO7 encodes importin-7, a karyopherin-β family nuclear transport receptor that mediates RanGTP-dependent import of cargo proteins through the nuclear pore complex. Importin-7 supports nuclear localization of diverse factors, including ribosomal proteins and regulatory proteins involved in transcriptional control, chromatin organization, and cell-cycle progression, thereby linking nucleocytoplasmic trafficking to proteostasis and growth signaling. Through its role in nuclear import, IPO7 influences ribosome biogenesis and stress-response programs that are frequently altered in proliferative states. Dysregulated nuclear transport and altered IPO7 expression have been associated with oncogenic pathways and genome stability phenotypes, making IPO7 a relevant node for mechanistic studies of nuclear trafficking in human cells.

Importin-7 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous IPO7 expression without altering the underlying DNA sequence.

Importin-7 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the IPO7 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 IPO7 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Importin-7 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native IPO7 locus and enabling the study of Importin-7-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Importin-7 pathway restoration in tumor cells with silenced or reduced IPO7 expression.

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