GCSH CRISPR/Cas9 KO Plasmid (h): sc-406784
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GCSH 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 GCSH 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
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
GCSH CRISPR/Cas9 KO Plasmid (h) | sc-406784 | 20 µg | $397.00 |
Overview
GCSH encodes the H protein of the glycine cleavage system, a mitochondrial lipoate-bearing carrier that shuttles reaction intermediates among glycine decarboxylase (P protein), aminomethyltransferase (T protein), and dihydrolipoamide dehydrogenase (L protein). Through this multienzyme complex, GCSH supports glycine catabolism and one-carbon metabolism by coupling glycine breakdown to folate-dependent carbon unit transfer, linking mitochondrial amino acid turnover to cellular redox and metabolic homeostasis. Perturbation of glycine cleavage activity is associated with altered mitochondrial function and imbalanced glycine/serine flux, processes relevant to neurometabolic dysregulation. As a node connecting lipoate-dependent enzymology with folate-mediated pathways, GCSH is frequently studied in the context of mitochondrial metabolism, oxidative stress responses, and inborn errors of glycine handling.
GCSH CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the GCSH gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the GCSH 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 GCSH 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 GCSH protein expression.
This CRISPR knockout system enables efficient generation of GCSH-deficient cell models for investigation of GCSH signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting GCSH exon(s) critical for GCSH 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 GCSH genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by GCSH CRISPR/Cas9 KO Plasmid (h) and GCSH CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the GCSH locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by GCSH HDR Plasmid (h) and GCSH HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by GCSH homology arms to support homology-directed repair at defined GCSH 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.