TID-1_L/S CRISPR Activation Plasmid (h): sc-404796-ACT

DNAJA3 encodes the mitochondrial DnaJ/Hsp40 co-chaperone TID-1, expressed as long and short isoforms (TID-1L/S) that coordinate Hsp70-dependent protein folding, import, and quality control within the mitochondrion. Through regulation of proteostasis and mitochondrial stress responses, TID-1L/S influences respiratory chain function, reactive oxygen species handling, and mitochondrial dynamics that shape cell survival and metabolic signaling. DNAJA3 has been linked to pathways governing apoptosis and cellular stress adaptation, positioning it as a useful node for studying mitochondrial dysfunction in cancer biology and neurodegeneration-associated phenotypes. Perturbation of TID-1L/S activity can alter mitochondrial homeostasis and downstream signaling programs relevant to proteotoxic stress and energy metabolism.

TID-1L/S CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DNAJA3 expression without altering the underlying DNA sequence.

TID-1L/S CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DNAJA3 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 DNAJA3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TID-1L/S expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DNAJA3 locus and enabling the study of TID-1L/S-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TID-1L/S pathway restoration in tumor cells with silenced or reduced DNAJA3 expression.

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