Ribosomal Protein L39 CRISPR Activation Plasmid (h): sc-410990-ACT

RPL39 encodes ribosomal protein L39, a small basic component of the 60S large ribosomal subunit that contributes to ribosome assembly and stable rRNA–protein interactions required for efficient cytosolic translation. As part of the core translational machinery, RPL39 influences proteostasis and cell growth programs that are tightly coupled to mTOR signaling, integrated stress responses, and ribosome biogenesis. Perturbations in ribosomal protein expression can shift translational capacity and fidelity, altering synthesis of regulators involved in cell-cycle control and stress adaptation. Dysregulated ribosome biogenesis and translation are recurrent features of multiple cancers and inherited ribosomopathies, making RPL39 a useful node for investigating how translational output shapes disease-relevant phenotypes.

Ribosomal Protein L39 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RPL39 expression without altering the underlying DNA sequence.

Ribosomal Protein L39 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RPL39 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 RPL39 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Ribosomal Protein L39 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RPL39 locus and enabling the study of Ribosomal Protein L39-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Ribosomal Protein L39 pathway restoration in tumor cells with silenced or reduced RPL39 expression.

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