Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
HIF PHD3 CRISPR/Cas9 KO Plasmid (h): sc-403671
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- HIF PHD3 CRISPR/Cas9 Knockout (KO) Plasmid (h) is a pool of plasmids, each encoding Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed for maximum knockout efficiency using sequences derived from the GeCKO v2 library
- gRNA sequences direct Cas9 to induce site-specific double-strand breaks (DSBs) in the HIF PHD3 genomic locus, resulting in gene knockout through non-homologous end joining (NHEJ)
- The puromycin resistance and RFP genes are flanked by LoxP sites, enabling removal of selection markers via Cre recombinase (Cre Vector: sc-418923) after establishing stable knockout cell lines
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: HIF PHD3 Antibody (EG188e/d5): sc-517601
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
HIF PHD3 CRISPR/Cas9 KO Plasmid (h) | sc-403671 | 20 µg | $397.00 |
Overview
EGLN3 encodes HIF prolyl hydroxylase 3 (PHD3), an oxygen-sensing dioxygenase that hydroxylates HIF-α subunits in an oxygen- and 2-oxoglutarate–dependent manner, promoting VHL-mediated ubiquitination and proteasomal turnover. Through this canonical hypoxia pathway, PHD3 helps tune transcriptional programs controlling angiogenesis, glycolytic metabolism, erythropoiesis, and cell survival in response to fluctuating oxygen tension. EGLN3 activity also intersects with mitochondrial metabolism and reactive oxygen species signaling, linking nutrient status and oxygen availability to adaptive gene expression. Dysregulation of EGLN3/PHD3 expression or function has been associated with altered hypoxic responses in cancer biology, ischemia-related cellular stress, and neuroendocrine tumor contexts, supporting its use as a mechanistic node in hypoxia research.
HIF PHD3 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the EGLN3 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the EGLN3 together with the Streptococcus pyogenes Cas9 nuclease. The plasmids also encode GFP, allowing fluorescent identification and enrichment of successfully transfected cells by fluorescence microscopy or flow cytometry.
The multi-guide design increases the likelihood of generating insertions or deletions (indels) that disrupt the EGLN3 open reading frame following Cas9-mediated double-strand break formation. DNA breaks introduced by the CRISPR/Cas9 system are repaired through endogenous non-homologous end joining (NHEJ) pathways, frequently resulting in frameshift mutations that abolish HIF PHD3 protein expression.
This CRISPR knockout system enables efficient generation of EGLN3-deficient cell models for investigation of HIF PHD3 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting EGLN3 exon(s) critical for HIF PHD3 function
- Co-expression of SpCas9 and sgRNA from a single plasmid for simplified delivery
- GFP reporter for identification of transfected cells
- Pool of plasmids targeting multiple EGLN3 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by HIF PHD3 CRISPR/Cas9 KO Plasmid (h) and HIF PHD3 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the EGLN3 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by HIF PHD3 HDR Plasmid (h) and HIF PHD3 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by EGLN3 homology arms to support homology-directed repair at defined EGLN3 target sites corresponding to the CRISPR/Cas9 KO designs. HDR donor availability may vary. See Related Products for availability.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.