MRP-S30 CRISPR Activation Plasmid (h): sc-411530-ACT

MRPS30 encodes mitochondrial ribosomal protein S30 (MRP-S30), a nuclear-encoded component of the 28S small subunit of the human mitoribosome required for mitochondrial mRNA translation. By supporting synthesis of oxidative phosphorylation (OXPHOS) subunits encoded by mitochondrial DNA, MRP-S30 contributes to respiratory chain assembly, ATP production, and mitochondrial proteostasis. Perturbation of mitoribosomal proteins can drive mitochondrial stress signaling, altered bioenergetics, and shifts in apoptosis susceptibility. MRPS30 expression changes have been reported across multiple disease contexts where mitochondrial metabolism is rewired, making it a useful node for studying mitonuclear communication and metabolic regulation.

MRP-S30 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MRPS30 expression without altering the underlying DNA sequence.

MRP-S30 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MRPS30 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 MRPS30 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MRP-S30 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MRPS30 locus and enabling the study of MRP-S30-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MRP-S30 pathway restoration in tumor cells with silenced or reduced MRPS30 expression.

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