Ribosomal Protein L3 CRISPR Activation Plasmid (h): sc-403160-ACT

RPL3 encodes ribosomal protein L3, an essential structural and functional component of the 60S large ribosomal subunit that contributes to ribosome biogenesis and peptidyl transferase center architecture during translation. As part of the core translational machinery, RPL3 supports global protein synthesis and is functionally connected to pathways governing nucleolar function, cellular growth control, and proteostasis. Perturbations in ribosomal protein homeostasis can engage ribosomal stress signaling, including p53-associated responses, and intersect with cell cycle regulation. Dysregulated expression of ribosomal genes, including RPL3, has been observed across multiple disease contexts and is frequently studied as a marker and modulator of altered translational capacity in proliferative states.

Ribosomal Protein L3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RPL3 expression without altering the underlying DNA sequence.

Ribosomal Protein L3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RPL3 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 RPL3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Ribosomal Protein L3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RPL3 locus and enabling the study of Ribosomal Protein L3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Ribosomal Protein L3 pathway restoration in tumor cells with silenced or reduced RPL3 expression.

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