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.
SphK1 CRISPR/Cas9 KO Plasmid (m): sc-423102
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- SphK1 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 SphK1 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: SphK1 Antibody (G-11): sc-365401
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
SphK1 CRISPR/Cas9 KO Plasmid (m) | sc-423102 | 20 µg | $397.00 |
Overview
Sphk1 encodes sphingosine kinase 1 (SphK1), a key lipid kinase that phosphorylates sphingosine to generate sphingosine-1-phosphate (S1P), thereby modulating the balance between pro-survival S1P and pro-apoptotic ceramide/sphingosine pools. SphK1-driven S1P signaling intersects with GPCR and receptor tyrosine kinase pathways to influence MAPK/ERK, PI3K/AKT, NF-κB, calcium flux, and cytoskeletal remodeling that collectively regulate proliferation, apoptosis resistance, migration, and inflammatory responses. In mouse systems, SphK1 is widely used to probe bioactive sphingolipid metabolism in immune regulation, vascular biology, and stress responses, including models of inflammation, fibrosis, and cancer-related phenotypes. Altered SPHK1/S1P axis activity is frequently linked to dysregulated cytokine production, barrier function, and tumor-associated signaling, making it a relevant node for mechanistic pathway studies.
SphK1 CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Sphk1 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Sphk1 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 Sphk1 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 SphK1 protein expression.
This CRISPR knockout system enables efficient generation of Sphk1-deficient cell models for investigation of SphK1 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Sphk1 exon(s) critical for SphK1 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 Sphk1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by SphK1 CRISPR/Cas9 KO Plasmid (m) and SphK1 CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Sphk1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by SphK1 HDR Plasmid (m) and SphK1 HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Sphk1 homology arms to support homology-directed repair at defined Sphk1 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.