Pol III RPC32 CRISPR Activation Plasmid (h): sc-417072-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Pol III RPC32 CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- Pol III RPC32 CRISPR Activation Plasmid (h) consists of three plasmids at a 1:1:1 mass ratio: a plasmid encoding the deactivated Cas9 (dCas9) nuclease (D10A and N863A) fused to the transactivation domain VP64, and a blasticidin resistance gene; a plasmid encoding the MS2-p65-HSF1 fusion protein, and a hygromycin resistance gene; a plasmid encoding a target-specific 20 nt guide RNA fused to two MS2 RNA aptamers, and a puromycin resistance gene
- The resulting SAM complex binds to a site-specific region approximately 200-250 nt upstream of the transcriptional start site and provides robust recruitment of transcription factors for highly efficient gene activation
- gRNAs encoded by Pol III RPC32 CRISPR Activation Plasmid (h) and Pol III RPC32 CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the POLR3G transcriptional start site. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Pol III RPC32 Antibody (H-9): sc-48365
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Pol III RPC32 CRISPR Activation Plasmid (h) | sc-417072-ACT | 20 µg | $397.00 | |||
Pol III RPC32 CRISPR Activation Plasmid (h2) | sc-417072-ACT-2 | 20 µg | $397.00 |
POLR3G encodes the Pol III subunit RPC32, a core component of RNA polymerase III required for transcription of small noncoding RNAs, including tRNAs and 5S rRNA, that support ribosome biogenesis and proteostasis. Through its role in Pol III assembly and promoter-dependent initiation, Pol III RPC32 contributes to cellular growth programs and metabolic adaptation linked to nutrient sensing and stress responses. Altered regulation of Pol III output and its accessory subunits has been associated with dysregulated proliferation and innate immune signaling pathways, making POLR3G a useful target for investigating transcriptional control of small RNA networks. Studying POLR3G function can help clarify how Pol III activity interfaces with chromatin regulation and RNA homeostasis in disease-relevant cellular states.
Pol III RPC32 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous POLR3G expression without altering the underlying DNA sequence.
Pol III RPC32 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the POLR3G 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 POLR3G transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Pol III RPC32 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native POLR3G locus and enabling the study of Pol III RPC32-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Pol III RPC32 pathway restoration in tumor cells with silenced or reduced POLR3G expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.