Haspin CRISPR Activation Plasmid (h): sc-403076-ACT

GSG2 encodes the serine/threonine kinase Haspin, a mitotic regulator that phosphorylates histone H3 at Thr3 to create a docking platform for the chromosomal passenger complex during prometaphase and metaphase. This epigenetic mark supports centromeric enrichment of Aurora B signaling and contributes to proper chromosome alignment, cohesion dynamics, and accurate segregation during cell division. Through its integration with mitotic checkpoint control and chromatin-based signaling, Haspin links histone phosphorylation to kinetochore function and spindle organization. Dysregulated Haspin activity has been associated with proliferative phenotypes and genomic instability, making GSG2 a useful node for studying cell-cycle–dependent chromatin regulation in cancer-relevant contexts.

Haspin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GSG2 expression without altering the underlying DNA sequence.

Haspin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GSG2 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 GSG2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Haspin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GSG2 locus and enabling the study of Haspin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Haspin pathway restoration in tumor cells with silenced or reduced GSG2 expression.

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