Ribosomal Protein L11 CRISPR Activation Plasmid (h): sc-401955-ACT

RPL11 encodes ribosomal protein L11, a core constituent of the 60S large ribosomal subunit that supports ribosome biogenesis and efficient translation elongation. Beyond its structural role, RPL11 participates in nucleolar stress signaling by modulating the MDM2–TP53 axis, linking disruptions in ribosome assembly to p53-dependent cell-cycle control and apoptosis. Altered RPL11 dosage or function is associated with ribosomopathy phenotypes and cancer-relevant pathways through effects on proteostasis, growth regulation, and checkpoint surveillance. As a result, RPL11 is frequently studied in contexts such as nucleolar integrity, translational control, and stress-responsive tumor suppressor networks.

Ribosomal Protein L11 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RPL11 expression without altering the underlying DNA sequence.

Ribosomal Protein L11 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RPL11 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 RPL11 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Ribosomal Protein L11 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RPL11 locus and enabling the study of Ribosomal Protein L11-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Ribosomal Protein L11 pathway restoration in tumor cells with silenced or reduced RPL11 expression.

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