MRP-S18C CRISPR Activation Plasmid (h): sc-412094-ACT

MRPS18C encodes the human mitochondrial ribosomal protein S18C (MRP-S18C), a component of the 28S small subunit of the mitoribosome required for translation of mitochondrially encoded oxidative phosphorylation (OXPHOS) polypeptides. By supporting synthesis of respiratory chain subunits, MRP-S18C helps maintain mitochondrial proteostasis, electron transport chain assembly, and ATP production, linking it to cellular energy homeostasis and mitochondrial stress responses. Perturbation of mitoribosomal function can impair mitochondrial translation and trigger integrated stress signaling, with downstream effects on redox balance and metabolic reprogramming. As a result, MRPS18C is relevant to studies of mitochondrial dysfunction and phenotypes observed in disorders featuring compromised bioenergetics.

MRP-S18C CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MRPS18C expression without altering the underlying DNA sequence.

MRP-S18C CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MRPS18C 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 MRPS18C transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MRP-S18C expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MRPS18C locus and enabling the study of MRP-S18C-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MRP-S18C pathway restoration in tumor cells with silenced or reduced MRPS18C expression.

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