NIRF CRISPR Activation Plasmid (h): sc-403908-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- NIRF CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- NIRF 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 NIRF CRISPR Activation Plasmid (h) and NIRF CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the UHRF2 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: NIRF Antibody (C-10): sc-398953
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
NIRF CRISPR Activation Plasmid (h) | sc-403908-ACT | 20 µg | $397.00 |
UHRF2 encodes NIRF, a multidomain nuclear protein that integrates E3 ubiquitin ligase activity with recognition of methylated DNA and histone marks to coordinate epigenetic regulation. Through its ubiquitin-like, PHD, and RING domains, NIRF contributes to chromatin remodeling, DNA methylation maintenance, and cell-cycle–linked control of transcriptional programs. These functions connect UHRF2 to pathways governing genome stability, DNA damage responses, and proliferation control, making it relevant to studies of aberrant epigenetic states in cancer and other disorders involving disrupted chromatin regulation. Altered UHRF2/NIRF activity has been associated with changes in promoter methylation patterns and transcriptional silencing or activation of genes implicated in oncogenic transformation and differentiation.
NIRF CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous UHRF2 expression without altering the underlying DNA sequence.
NIRF CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the UHRF2 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 UHRF2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NIRF expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native UHRF2 locus and enabling the study of NIRF-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NIRF pathway restoration in tumor cells with silenced or reduced UHRF2 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.