MRP-S18B CRISPR Activation Plasmid (h): sc-412012-ACT

MRPS18B encodes the human mitochondrial ribosomal protein MRP-S18B, a component of the 28S small subunit that supports translation of mitochondrially encoded oxidative phosphorylation proteins. By contributing to mitochondrial protein synthesis, MRP-S18B helps maintain electron transport chain integrity, ATP production, and overall mitochondrial homeostasis. Perturbation of mitoribosome function can remodel cellular energy metabolism and elevate oxidative stress, linking MRPS18B-associated biology to pathways governing mitochondrial quality control, proteostasis, and stress signaling. Altered mitochondrial translation capacity is frequently observed in contexts of neurodegeneration, cardiometabolic dysfunction, and cancer-associated metabolic rewiring, making MRPS18B a useful node for mechanistic studies.

MRP-S18B CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MRPS18B expression without altering the underlying DNA sequence.

MRP-S18B CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MRPS18B 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 MRPS18B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MRP-S18B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MRPS18B locus and enabling the study of MRP-S18B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MRP-S18B pathway restoration in tumor cells with silenced or reduced MRPS18B expression.

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