TPX2 CRISPR Activation Plasmid (h): sc-402581-ACT

TPX2 (targeting protein for Xklp2) is a microtubule-associated spindle assembly factor that coordinates centrosome-independent microtubule nucleation and stabilizes kinetochore fibers during mitosis. It functions in the Ran-GTP/importin-regulated pathway to spatially activate Aurora A kinase and organize bipolar spindle architecture, supporting faithful chromosome congression and segregation. Dysregulated TPX2 expression perturbs mitotic checkpoint control, promotes chromosomal instability, and is frequently associated with proliferative phenotypes observed across diverse tumor contexts. As a result, TPX2 is widely used as a mechanistic node for studying cell cycle progression, spindle dynamics, and genome maintenance in human cell models.

TPX2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TPX2 expression without altering the underlying DNA sequence.

TPX2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TPX2 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 TPX2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TPX2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TPX2 locus and enabling the study of TPX2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TPX2 pathway restoration in tumor cells with silenced or reduced TPX2 expression.

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