Ribosomal Protein S14 CRISPR Activation Plasmid (h): sc-403429-ACT

RPS14 encodes ribosomal protein S14, an essential component of the 40S small ribosomal subunit that supports accurate mRNA decoding and initiation of protein synthesis. Through its role in ribosome biogenesis and translational control, RPS14 contributes to proteostasis and cell-cycle progression, linking it to stress-response programs such as nucleolar surveillance and p53-associated checkpoints. Altered ribosomal protein function can perturb global translation and selectively affect transcripts involved in growth and differentiation, making RPS14 a useful node for studying ribosomopathies and proliferative phenotypes. Dysregulation of ribosome assembly factors, including RPS14, has been associated with hematopoietic defects and cancer-related translational remodeling in experimental systems.

Ribosomal Protein S14 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RPS14 expression without altering the underlying DNA sequence.

Ribosomal Protein S14 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RPS14 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 RPS14 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Ribosomal Protein S14 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RPS14 locus and enabling the study of Ribosomal Protein S14-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Ribosomal Protein S14 pathway restoration in tumor cells with silenced or reduced RPS14 expression.

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