KIF14 CRISPR Activation Plasmid (h): sc-408805-ACT

KIF14 encodes a kinesin motor protein that localizes to the mitotic spindle and midbody, where it supports chromosome segregation, spindle organization, and cytokinesis. It coordinates microtubule-based transport and interfaces with central spindle regulators to ensure proper abscission and cell-cycle progression. Dysregulated KIF14 expression is frequently associated with proliferative phenotypes and chromosomal instability, linking it to pathways governing mitotic fidelity and genome maintenance. As a result, KIF14 is commonly studied in models of cell division control, aneuploidy, and tumor biology to understand how altered kinesin activity reshapes mitotic outcomes.

KIF14 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KIF14 expression without altering the underlying DNA sequence.

KIF14 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KIF14 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 KIF14 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous KIF14 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KIF14 locus and enabling the study of KIF14-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of KIF14 pathway restoration in tumor cells with silenced or reduced KIF14 expression.

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