POLR3GL CRISPR Activation Plasmid (h): sc-413599-ACT

Human POLR3GL encodes a small subunit of RNA polymerase III that supports transcription of short noncoding RNAs, including tRNAs, 5S rRNA, and other Pol III-dependent RNAs essential for translation capacity and cellular homeostasis. POLR3GL contributes to Pol III complex assembly and promoter-dependent initiation, linking its activity to growth control, stress adaptation, and innate immune signaling pathways influenced by Pol III-derived RNAs. Dysregulation of Pol III transcriptional programs is broadly relevant to proliferative and developmental phenotypes, making POLR3GL a useful node for studying how altered noncoding RNA output reshapes gene expression networks. Investigating POLR3GL function can inform mechanisms connecting Pol III activity to transcriptional regulation, protein synthesis demands, and disease-associated cellular states.

POLR3GL CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous POLR3GL expression without altering the underlying DNA sequence.

POLR3GL CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the POLR3GL 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 POLR3GL transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous POLR3GL expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native POLR3GL locus and enabling the study of POLR3GL-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of POLR3GL pathway restoration in tumor cells with silenced or reduced POLR3GL expression.

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