TARDBP CRISPR Activation Plasmid (h): sc-401890-ACT

TARDBP encodes TDP-43, an RNA/DNA-binding protein that regulates pre-mRNA splicing, mRNA stability, and transport, with prominent roles in neuronal RNA metabolism and stress granule dynamics. Through binding UG-rich RNA motifs, TDP-43 modulates alternative splicing programs and maintains transcriptome integrity, linking it to pathways that control proteostasis, nucleocytoplasmic transport, and cellular stress responses. Dysregulation of TDP-43 localization and aggregation is a hallmark of multiple neurodegenerative disorders, and altered TARDBP-dependent splicing has been implicated in disease-relevant gene networks. As a result, TARDBP is widely studied for mechanisms of RNA processing, neurotoxicity, and gene expression control in human cell models.

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

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

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