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.
GALK1 CRISPR/Cas9 KO Plasmid (h): sc-405217
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GALK1 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 GALK1 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: GALK1 Antibody (A-2): sc-393404
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
GALK1 CRISPR/Cas9 KO Plasmid (h) | sc-405217 | 20 µg | $397.00 |
Overview
GALK1 encodes galactokinase 1, a cytosolic enzyme that catalyzes phosphorylation of α-D-galactose to galactose-1-phosphate, initiating flux through the Leloir pathway for galactose utilization. By controlling entry of galactose into glycolytic and glycoconjugate biosynthesis routes, GALK1 activity contributes to cellular energy metabolism and glycan homeostasis. Disruption of galactose handling is linked to inborn errors of metabolism, and altered Leloir pathway function can influence oxidative stress responses and metabolite toxicity in diverse cellular contexts. GALK1 is therefore relevant for studying carbohydrate metabolism, metabolic rewiring, and downstream effects on protein and lipid glycosylation.
GALK1 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the GALK1 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the GALK1 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 GALK1 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 GALK1 protein expression.
This CRISPR knockout system enables efficient generation of GALK1-deficient cell models for investigation of GALK1 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting GALK1 exon(s) critical for GALK1 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 GALK1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by GALK1 CRISPR/Cas9 KO Plasmid (h) and GALK1 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the GALK1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by GALK1 HDR Plasmid (h) and GALK1 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by GALK1 homology arms to support homology-directed repair at defined GALK1 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.