MFG-E8 CRISPR/Cas9 KO Plasmid (m): sc-421639
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- MFG-E8 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 MFG-E8 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: MFG-E8 Antibody (H-3): sc-377356
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
MFG-E8 CRISPR/Cas9 KO Plasmid (m) | sc-421639 | 20 µg | $397.00 |
Overview
Mfge8 encodes milk fat globule-EGF factor 8 (MFG-E8), a secreted glycoprotein that bridges phosphatidylserine on apoptotic cells to integrins such as ITGAV/ITGB3 and ITGAV/ITGB5 on phagocytes, promoting efferocytosis and limiting secondary necrosis. Through this opsonization-like activity, MFG-E8 influences immune homeostasis, resolution of inflammation, and tissue remodeling, with downstream effects on cytokine signaling and macrophage activation states. In mouse systems, altered Mfge8 function has been linked to dysregulated clearance of dying cells, chronic inflammatory phenotypes, and impaired repair responses across vascular, metabolic, and neuroinflammatory contexts.
MFG-E8 CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Mfge8 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Mfge8 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 Mfge8 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 MFG-E8 protein expression.
This CRISPR knockout system enables efficient generation of Mfge8-deficient cell models for investigation of MFG-E8 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Mfge8 exon(s) critical for MFG-E8 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 Mfge8 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by MFG-E8 CRISPR/Cas9 KO Plasmid (m) and MFG-E8 CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Mfge8 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by MFG-E8 HDR Plasmid (m) and MFG-E8 HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Mfge8 homology arms to support homology-directed repair at defined Mfge8 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.