TARSL1 CRISPR Activation Plasmid (h): sc-410152-ACT

Human TARS2 encodes a mitochondrial threonyl-tRNA synthetase required for charging mt-tRNA(Thr) and maintaining fidelity of mitochondrial translation. By supporting synthesis of oxidative phosphorylation (OXPHOS) subunits, TARSL1/TARS2 activity is tightly linked to mitochondrial proteostasis, respiratory chain function, and cellular energy metabolism. Perturbation of this axis can remodel integrated stress signaling and mitochondrial-nuclear communication pathways that influence proliferation and survival. Dysregulated mitochondrial translation and OXPHOS are recurrent features across neurodevelopmental and neuromuscular phenotypes as well as cancer metabolism, making TARS2 a useful node for mechanistic studies of mitochondrial dysfunction.

TARSL1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TARS2 expression without altering the underlying DNA sequence.

TARSL1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TARS2 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 TARS2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TARSL1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TARS2 locus and enabling the study of TARSL1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TARSL1 pathway restoration in tumor cells with silenced or reduced TARS2 expression.

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