HIF PHD2 CRISPR Activation Plasmid (h): sc-403334-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- HIF PHD2 CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- HIF PHD2 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 HIF PHD2 CRISPR Activation Plasmid (h) and HIF PHD2 CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the EGLN1 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: HIF PHD2 Antibody (H-8): sc-271835
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
HIF PHD2 CRISPR Activation Plasmid (h) | sc-403334-ACT | 20 µg | $397.00 |
EGLN1 encodes HIF PHD2, a 2-oxoglutarate/Fe(II)-dependent prolyl hydroxylase that hydroxylates HIF-α subunits under normoxia, enabling von Hippel–Lindau (VHL) recognition and proteasomal turnover. Through this oxygen-sensing pathway, HIF PHD2 modulates transcriptional programs controlling angiogenesis, erythropoiesis, glycolytic metabolism, mitochondrial adaptation, and cell survival during hypoxic stress. Altered EGLN1/PHD2 activity impacts HIF signaling amplitude and has been linked to hypoxia adaptation phenotypes and diseases characterized by dysregulated oxygen homeostasis, including cancer biology, pulmonary vascular remodeling, and erythrocytosis-related mechanisms. As a central node in cellular oxygen sensing, EGLN1 is widely studied for its effects on hypoxia-inducible gene networks and metabolic reprogramming.
HIF PHD2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EGLN1 expression without altering the underlying DNA sequence.
HIF PHD2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EGLN1 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 EGLN1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HIF PHD2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EGLN1 locus and enabling the study of HIF PHD2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HIF PHD2 pathway restoration in tumor cells with silenced or reduced EGLN1 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.