GTPBP10 CRISPR Activation Plasmid (h): sc-413794-ACT

Human GTPBP10 encodes a mitochondrial GTP-binding protein implicated in late-stage mitoribosome maturation, supporting assembly of the large ribosomal subunit and efficient mitochondrial translation. By regulating production of oxidative phosphorylation components encoded by mitochondrial DNA, GTPBP10 contributes to respiratory chain integrity, ATP generation, and mitochondrial proteostasis. Perturbation of mitoribosome biogenesis and mitochondrial translation is broadly linked to bioenergetic stress responses, altered redox balance, and mitochondrial dysfunction pathways that are frequently investigated in neuromuscular and metabolic disease models as well as cancer cell adaptation. As a result, GTPBP10 serves as a useful node for studying mitochondrial ribosome assembly, OXPHOS regulation, and downstream stress signaling.

GTPBP10 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GTPBP10 expression without altering the underlying DNA sequence.

GTPBP10 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GTPBP10 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 GTPBP10 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GTPBP10 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GTPBP10 locus and enabling the study of GTPBP10-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GTPBP10 pathway restoration in tumor cells with silenced or reduced GTPBP10 expression.

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