Ribosomal Protein L18 CRISPR Activation Plasmid (h): sc-404259-ACT

RPL18 encodes ribosomal protein L18, a core component of the 60S large ribosomal subunit that supports rRNA binding and assembly of translation-competent ribosomes. By contributing to ribosome biogenesis and mRNA translation, RPL18 influences proteostasis programs that interface with cell growth control, stress adaptation, and checkpoint signaling. Altered expression of ribosomal proteins and nucleolar stress responses are frequently linked to aberrant proliferation and RNA metabolism phenotypes, making RPL18 a useful node for studying translational regulation in disease-relevant contexts. Perturbation of ribosome assembly factors can also impact p53-associated surveillance pathways through disrupted ribosomal homeostasis.

Ribosomal Protein L18 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RPL18 expression without altering the underlying DNA sequence.

Ribosomal Protein L18 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RPL18 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 RPL18 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Ribosomal Protein L18 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RPL18 locus and enabling the study of Ribosomal Protein L18-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Ribosomal Protein L18 pathway restoration in tumor cells with silenced or reduced RPL18 expression.

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