POLR1E CRISPR Activation Plasmid (h): sc-407602-ACT

POLR1E encodes a core subunit of RNA polymerase I, a multiprotein complex responsible for transcribing ribosomal RNA precursors that initiate ribosome biogenesis in the nucleolus. By supporting pre-rRNA synthesis and processing, POLR1E contributes to cellular growth control, proteostasis capacity, and nucleolar stress signaling pathways that link ribosome production to cell cycle regulation. Perturbation of RNA polymerase I function can disrupt protein synthesis homeostasis and activate stress responses such as p53-associated checkpoints, making POLR1E relevant for studies of proliferation, differentiation, and genome stability. Genetic evidence connects POLR1E dysfunction to human ribosomopathy phenotypes and developmental disorders, underscoring its importance in tissues with high biosynthetic demand.

POLR1E CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous POLR1E expression without altering the underlying DNA sequence.

POLR1E CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the POLR1E 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 POLR1E transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous POLR1E expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native POLR1E locus and enabling the study of POLR1E-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of POLR1E pathway restoration in tumor cells with silenced or reduced POLR1E expression.

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