HRI CRISPR/Cas9 KO Plasmid (m): sc-420950
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- HRI CRISPR/Cas9 Knockout (KO) Plasmid (m) 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 HRI 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: HRI Antibody (D-12): sc-365239
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
HRI CRISPR/Cas9 KO Plasmid (m) | sc-420950 | 20 µg | $397.00 |
Overview
Eif2ak1 encodes heme-regulated inhibitor (HRI), a stress-responsive eIF2α kinase that phosphorylates EIF2S1 to attenuate global translation while promoting selective translation of stress-adaptive transcripts. In mouse cells, HRI links heme availability, oxidative stress, and proteotoxic stress to the integrated stress response, coordinating proteostasis, redox balance, and mitochondrial homeostasis. This signaling axis interfaces with pathways controlling heme biosynthesis and iron metabolism, and modulates inflammatory and apoptotic programs under cellular stress. Dysregulation of HRI-dependent translational control is relevant to models of anemia and hemoglobinopathies, neurodegeneration, and stress-driven metabolic or inflammatory phenotypes where protein synthesis homeostasis is perturbed.
HRI CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Eif2ak1 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Eif2ak1 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 Eif2ak1 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 HRI protein expression.
This CRISPR knockout system enables efficient generation of Eif2ak1-deficient cell models for investigation of HRI signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Eif2ak1 exon(s) critical for HRI 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 Eif2ak1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by HRI CRISPR/Cas9 KO Plasmid (m) and HRI CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Eif2ak1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by HRI HDR Plasmid (m) and HRI HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Eif2ak1 homology arms to support homology-directed repair at defined Eif2ak1 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.