Ribosomal Protein L13A CRISPR Activation Plasmid (h): sc-403494-ACT

RPL13A encodes ribosomal protein L13A, a core component of the 60S large ribosomal subunit required for ribosome assembly and efficient translation elongation. Beyond its structural role in the ribosome, L13A participates in translational control programs linked to cellular stress and inflammatory signaling, including interferon-responsive pathways that modulate selective mRNA translation. Altered ribosome biogenesis and dysregulated protein synthesis are common features of proliferative and stress-adaptation states, making RPL13A relevant to studies of cell growth control and proteostasis. As a widely expressed ribosomal gene, RPL13A is also frequently used as a reference in gene expression workflows, and its regulation can inform interpretations of global translational remodeling.

Ribosomal Protein L13A CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RPL13A expression without altering the underlying DNA sequence.

Ribosomal Protein L13A CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RPL13A 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 RPL13A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Ribosomal Protein L13A expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RPL13A locus and enabling the study of Ribosomal Protein L13A-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Ribosomal Protein L13A pathway restoration in tumor cells with silenced or reduced RPL13A expression.

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