Ribosomal Protein S16 CRISPR Activation Plasmid (h): sc-403364-ACT

RPS16 encodes human ribosomal protein S16, a conserved component of the 40S small ribosomal subunit required for accurate mRNA decoding and efficient translation initiation. As part of ribosome biogenesis and cytoplasmic protein synthesis, RPS16 supports cellular growth programs and proteostasis pathways that couple nutrient signaling to translational output. Perturbations in ribosomal protein dosage and assembly can trigger nucleolar stress responses and reshape p53-linked transcriptional networks, connecting ribosome integrity to cell-cycle control. Aberrant expression of ribosomal proteins, including RPS16, has been reported in multiple disease contexts and is frequently examined as a marker of altered translational capacity in proliferative and stress-adapted states.

Ribosomal Protein S16 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RPS16 expression without altering the underlying DNA sequence.

Ribosomal Protein S16 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RPS16 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 RPS16 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Ribosomal Protein S16 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RPS16 locus and enabling the study of Ribosomal Protein S16-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Ribosomal Protein S16 pathway restoration in tumor cells with silenced or reduced RPS16 expression.

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