NPAT CRISPR Activation Plasmid (m): sc-434105-ACT
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- NPAT CRISPR Activation Plasmid (m) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- NPAT 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 NPAT CRISPR Activation Plasmid (m) and NPAT CRISPR Activation Plasmid (m2) target distinct regulatory regions upstream of the Npat transcriptional start site. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
NPAT CRISPR Activation Plasmid (m) | sc-434105-ACT | 20 µg | $397.00 | |||
NPAT CRISPR Activation Plasmid (m2) | sc-434105-ACT-2 | 20 µg | $397.00 |
Mouse Npat encodes NPAT, a nuclear cofactor that links cell-cycle progression to histone gene transcription and chromatin dynamics. NPAT is activated downstream of cyclin E/CDK2 and localizes to histone locus bodies, where it supports S-phase–coupled expression of replication-dependent histones and coordinates DNA replication and genome maintenance. Through these roles, NPAT contributes to pathways governing G1/S transition, nucleotide incorporation, and chromatin assembly, processes commonly perturbed in proliferative stress and disease-associated cell state changes. Modulating Npat expression is therefore useful for dissecting how cell-cycle signaling interfaces with transcriptional programs that control genome stability.
NPAT CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Npat expression without altering the underlying DNA sequence.
NPAT CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Npat 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 Npat transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NPAT expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Npat locus and enabling the study of NPAT-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NPAT pathway restoration in tumor cells with silenced or reduced Npat expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.