GCH-I CRISPR/Cas9 KO Plasmid (m): sc-420509
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GCH-I 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 GCH-I 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: GCH-I Antibody (C-4): sc-271482
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
GCH-I CRISPR/Cas9 KO Plasmid (m) | sc-420509 | 20 µg | $397.00 |
Overview
Mouse Gch1 encodes GTP cyclohydrolase I (GCH-I), the rate-limiting enzyme in tetrahydrobiopterin (BH4) biosynthesis from GTP. BH4 is an essential redox-active cofactor for aromatic amino acid hydroxylases involved in catecholamine and serotonin production, and for nitric oxide synthases that regulate nitric oxide signaling. Through its control of BH4 availability, GCH-I links pteridine metabolism to neurotransmitter homeostasis, vascular biology, and oxidative stress sensitivity. Altered GCH-I/BH4 axis activity has been associated with dysregulated monoamine pathways and impaired nitric oxide coupling in models of neurologic and cardiovascular dysfunction.
GCH-I CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Gch1 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Gch1 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 Gch1 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 GCH-I protein expression.
This CRISPR knockout system enables efficient generation of Gch1-deficient cell models for investigation of GCH-I signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Gch1 exon(s) critical for GCH-I 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 Gch1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by GCH-I CRISPR/Cas9 KO Plasmid (m) and GCH-I CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Gch1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by GCH-I HDR Plasmid (m) and GCH-I HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Gch1 homology arms to support homology-directed repair at defined Gch1 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.