Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
MtRPOL CRISPR Activation Plasmid (h): sc-403497-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- MtRPOL CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- MtRPOL 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 MtRPOL CRISPR Activation Plasmid (h) and MtRPOL CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the POLRMT 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: MtRPOL Antibody (B-1): sc-365082
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
MtRPOL CRISPR Activation Plasmid (h) | sc-403497-ACT | 20 µg | $397.00 |
Human POLRMT encodes the mitochondrial RNA polymerase (MtRPOL), the nuclear-encoded enzyme responsible for transcribing the mitochondrial genome to produce rRNAs, tRNAs, and mRNAs required for oxidative phosphorylation. MtRPOL functions within the mitochondrial transcription apparatus alongside TFAM and TFB2M and is tightly coupled to mitochondrial DNA maintenance, ribosome biogenesis, and respiratory chain assembly. By controlling mitochondrial gene expression, POLRMT supports ATP production, redox homeostasis, and stress responses linked to mitochondrial quality control pathways. Dysregulation of mitochondrial transcription and POLRMT activity is associated with impaired bioenergetics and is relevant to the study of mitochondrial disorders and neurodegeneration-associated mitochondrial dysfunction.
MtRPOL CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous POLRMT expression without altering the underlying DNA sequence.
MtRPOL CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the POLRMT 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 POLRMT transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MtRPOL expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native POLRMT locus and enabling the study of MtRPOL-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MtRPOL pathway restoration in tumor cells with silenced or reduced POLRMT expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.