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.
Ferrochelatase CRISPR/Cas9 KO Plasmid (h): sc-402043
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Ferrochelatase 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 Ferrochelatase 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: Ferrochelatase Antibody (A-3): sc-377377
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Ferrochelatase CRISPR/Cas9 KO Plasmid (h) | sc-402043 | 20 µg | $397.00 |
Overview
FECH encodes human ferrochelatase, a mitochondrial inner-membrane enzyme that catalyzes insertion of ferrous iron into protoporphyrin IX to produce heme, the terminal step of the porphyrin/heme biosynthesis pathway. By controlling heme availability, ferrochelatase influences erythroid maturation, mitochondrial respiration, redox balance, and hemoprotein assembly. Disruption of FECH activity is linked to porphyrin accumulation and altered heme homeostasis, with relevance to erythropoietic protoporphyria and broader metabolic stress phenotypes in cells with high heme demand. FECH is therefore widely studied in mitochondrial biology, iron handling, and porphyrin metabolism, including pathway responses to oxidative and metabolic perturbations.
Ferrochelatase CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the FECH gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the FECH 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 FECH 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 Ferrochelatase protein expression.
This CRISPR knockout system enables efficient generation of FECH-deficient cell models for investigation of Ferrochelatase signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting FECH exon(s) critical for Ferrochelatase 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 FECH genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Ferrochelatase CRISPR/Cas9 KO Plasmid (h) and Ferrochelatase CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the FECH locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Ferrochelatase HDR Plasmid (h) and Ferrochelatase HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by FECH homology arms to support homology-directed repair at defined FECH 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.