Ribosomal Protein L22L1 CRISPR Activation Plasmid (h): sc-402323-ACT

RPL22L1 encodes ribosomal protein L22-like 1, a component associated with the 60S large ribosomal subunit that supports ribosome biogenesis and efficient mRNA translation. As part of core translational machinery, RPL22L1 contributes to proteostasis and growth-related programs that couple nutrient sensing, cell-cycle progression, and stress responses through global and transcript-selective translational control. Variation in ribosomal protein dosage can influence ribosomal assembly and specialized translation, with downstream effects on pathways linked to proliferation and cellular adaptation. Dysregulated expression of ribosome-associated genes, including RPL22L1, is frequently investigated in contexts such as oncogenic growth, RNA metabolism alterations, and ribosomopathy-like phenotypes.

Ribosomal Protein L22L1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RPL22L1 expression without altering the underlying DNA sequence.

Ribosomal Protein L22L1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RPL22L1 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 RPL22L1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Ribosomal Protein L22L1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RPL22L1 locus and enabling the study of Ribosomal Protein L22L1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Ribosomal Protein L22L1 pathway restoration in tumor cells with silenced or reduced RPL22L1 expression.

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