TDP2 CRISPR Activation Plasmid (h): sc-403902-ACT

Human TDP2 (tyrosyl-DNA phosphodiesterase 2) is a DNA repair enzyme that removes 5′-phosphotyrosyl adducts generated by abortive topoisomerase II activity, thereby restoring ligatable DNA ends after double-strand break formation. This activity supports genome stability and cell survival following replication stress and genotoxic exposure, functioning within the broader double-strand break repair network coordinated with non-homologous end joining and related end-processing factors. TDP2 also contributes to RNA virus replication through roles linked to VPg unlinking in some viral systems, highlighting its relevance to host–pathogen interactions. Altered TDP2 function has been associated with neurodevelopmental phenotypes and DNA repair deficiency syndromes, making it a useful node for mechanistic studies of genome maintenance.

TDP2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TDP2 expression without altering the underlying DNA sequence.

TDP2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TDP2 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 TDP2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TDP2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TDP2 locus and enabling the study of TDP2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TDP2 pathway restoration in tumor cells with silenced or reduced TDP2 expression.

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