POLR2J1 CRISPR Activation Plasmid (h): sc-409187-ACT

POLR2J encodes a subunit of RNA polymerase II, the central enzyme responsible for synthesizing mRNA and many noncoding RNAs in human cells. POLR2J1 contributes to assembly and function of the Pol II complex and supports transcription initiation and elongation programs that shape global gene expression, RNA processing, and cellular responses to signaling cues. Through its role in basal transcription machinery, POLR2J1 is relevant to pathways controlling proliferation, stress adaptation, and differentiation states that depend on tightly regulated transcriptional output. Dysregulation of Pol II–associated factors is frequently observed in cancer and other disorders of gene expression control, making POLR2J1 a useful node for mechanistic studies of transcriptional homeostasis.

POLR2J1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous POLR2J expression without altering the underlying DNA sequence.

POLR2J1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the POLR2J 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 POLR2J transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous POLR2J1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native POLR2J locus and enabling the study of POLR2J1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of POLR2J1 pathway restoration in tumor cells with silenced or reduced POLR2J expression.

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