TRIP13 CRISPR Activation Plasmid (h): sc-404006-ACT

TRIP13 (thyroid hormone receptor interactor 13) encodes an AAA+ ATPase that regulates chromosome dynamics during mitosis and meiosis, including spindle checkpoint signaling and proper kinetochore–microtubule attachment. It functions in pathways controlling cell-cycle progression, error correction, and genome stability by remodeling key checkpoint components to ensure accurate chromosomal segregation. Dysregulated TRIP13 activity has been linked to aneuploidy and chromosomal instability phenotypes observed in multiple disease contexts, making it a useful node for studying checkpoint adaptation and proliferative stress. In human cell models, TRIP13 perturbation provides a tractable entry point for investigating mechanisms connecting mitotic surveillance to DNA damage responses and replication-associated instability.

TRIP13 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TRIP13 expression without altering the underlying DNA sequence.

TRIP13 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TRIP13 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 TRIP13 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TRIP13 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TRIP13 locus and enabling the study of TRIP13-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TRIP13 pathway restoration in tumor cells with silenced or reduced TRIP13 expression.

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