RPL29 CRISPR Activation Plasmid (h): sc-403879-ACT

RPL29 encodes ribosomal protein L29, a basic component of the 60S large ribosomal subunit that contributes to ribosome assembly and efficient translation of mRNAs in the cytoplasm. As part of the core protein synthesis machinery, RPL29 supports proteostasis and cellular growth programs that are tightly coupled to ribosome biogenesis and translational control. Altered expression of ribosomal proteins, including RPL29, is frequently observed in conditions featuring dysregulated proliferation and stress adaptation, and can influence cellular phenotypes through changes in global and selective translation. RPL29 therefore serves as a useful node for studying how ribosome composition and translational capacity interface with cell cycle control, metabolic rewiring, and stress-response pathways.

RPL29 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RPL29 expression without altering the underlying DNA sequence.

RPL29 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RPL29 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 RPL29 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous RPL29 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RPL29 locus and enabling the study of RPL29-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of RPL29 pathway restoration in tumor cells with silenced or reduced RPL29 expression.

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