Ribosomal Protein S5 CRISPR Activation Plasmid (h): sc-404960-ACT

Human RPS5 encodes ribosomal protein S5, a core component of the 40S small ribosomal subunit required for accurate mRNA decoding and translation initiation. As part of cytosolic ribosome biogenesis, RPS5 supports rRNA processing, assembly of pre-40S particles, and maintenance of translational fidelity that influences global protein homeostasis. Perturbation of ribosomal protein dosage can trigger ribosome stress responses with downstream effects on cell-cycle control and apoptosis, linking altered ribosome function to oncogenic growth programs and congenital ribosomopathies. Because translational output shapes stress adaptation and proliferation, RPS5 is frequently studied in pathways connecting protein synthesis, proteostasis, and cellular fitness.

Ribosomal Protein S5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RPS5 expression without altering the underlying DNA sequence.

Ribosomal Protein S5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RPS5 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 RPS5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Ribosomal Protein S5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RPS5 locus and enabling the study of Ribosomal Protein S5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Ribosomal Protein S5 pathway restoration in tumor cells with silenced or reduced RPS5 expression.

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