DNA pol ζ CRISPR Activation Plasmid (m): sc-422655-ACT
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- DNA pol ζ CRISPR Activation Plasmid (m) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- DNA pol ζ CRISPR Activation Plasmid (m) 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 DNA pol ζ CRISPR Activation Plasmid (m) and DNA pol ζ CRISPR Activation Plasmid (m2) target distinct regulatory regions upstream of the Rev3l transcriptional start site. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
DNA pol ζ CRISPR Activation Plasmid (m) | sc-422655-ACT | 20 µg | $397.00 | |||
DNA pol ζ CRISPR Activation Plasmid (m2) | sc-422655-ACT-2 | 20 µg | $397.00 |
Rev3l encodes the catalytic subunit of DNA polymerase ζ (pol ζ), a specialized B-family polymerase essential for translesion DNA synthesis and tolerance of replication-blocking lesions. In mouse cells, pol ζ cooperates with REV1 and ubiquitin-modified PCNA within the DNA damage response to extend from mismatched or damaged primer termini, promoting replication fork progression under genotoxic stress. This activity influences mutagenesis rates and genome stability, linking Rev3l-regulated pathways to mechanisms that shape mutation accumulation during carcinogenesis and to cellular sensitivity to DNA-damaging agents. Because Rev3l function intersects with replication stress signaling and repair pathway choice, it is widely studied in models of genomic instability, immune diversification, and developmental viability.
DNA pol ζ CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Rev3l expression without altering the underlying DNA sequence.
DNA pol ζ CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Rev3l 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 Rev3l transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DNA pol ζ expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Rev3l locus and enabling the study of DNA pol ζ-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DNA pol ζ pathway restoration in tumor cells with silenced or reduced Rev3l expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.