DNA-PKCS CRISPR Activation Plasmid (m): sc-422405-ACT
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- DNA-PKCS CRISPR Activation Plasmid (m) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- DNA-PKCS 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-PKCS CRISPR Activation Plasmid (m) and DNA-PKCS CRISPR Activation Plasmid (m2) target distinct regulatory regions upstream of the Prkdc 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: DNA-PKCS Antibody (G-12): sc-390849
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
DNA-PKCS CRISPR Activation Plasmid (m) | sc-422405-ACT | 20 µg | $397.00 |
Prkdc encodes the catalytic subunit of DNA-dependent protein kinase, DNA-PKCS, a central regulator of DNA double-strand break repair by non-homologous end joining (NHEJ). DNA-PKCS forms an active holoenzyme with the Ku70/Ku80 complex to coordinate end recognition, synapsis, end processing, and ligation, and it interfaces with ATM/ATR signaling to shape checkpoint responses following genotoxic stress. In mouse cells, Prkdc activity is also essential for V(D)J recombination during lymphocyte development, linking this kinase to genome stability and immune system maturation. Perturbation of DNA-PKCS-dependent repair pathways is widely used to model radiosensitivity, chromosomal instability, and DNA repair defects relevant to cancer biology and immunodeficiency research.
DNA-PKCS CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Prkdc expression without altering the underlying DNA sequence.
DNA-PKCS CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Prkdc 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 Prkdc transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DNA-PKCS expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Prkdc locus and enabling the study of DNA-PKCS-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DNA-PKCS pathway restoration in tumor cells with silenced or reduced Prkdc expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.