Ribosomal Protein L28 CRISPR Activation Plasmid (h): sc-405270-ACT

RPL28 encodes ribosomal protein L28, a core component of the 60S large ribosomal subunit required for accurate translation elongation and efficient ribosome biogenesis. As part of the nucleolar ribosome assembly pathway, RPL28 supports rRNA processing, large subunit maturation, and proteostasis, linking its activity to cellular growth control and stress-adaptive translational reprogramming. Altered expression or dosage of ribosomal proteins can perturb global and selective mRNA translation and activate nucleolar stress signaling, processes frequently investigated in cancer biology and other disorders involving dysregulated protein synthesis. RPL28 is therefore a useful entry point for studying ribosome function, translational control networks, and downstream effects on cell cycle progression and metabolic adaptation.

Ribosomal Protein L28 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RPL28 expression without altering the underlying DNA sequence.

Ribosomal Protein L28 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RPL28 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 RPL28 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Ribosomal Protein L28 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RPL28 locus and enabling the study of Ribosomal Protein L28-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Ribosomal Protein L28 pathway restoration in tumor cells with silenced or reduced RPL28 expression.

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