MLTK CRISPR/Cas9 KO Plasmid (m): sc-437372
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- MLTK 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 MLTK 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: MLTK Antibody (A-2): sc-390924
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
MLTK CRISPR/Cas9 KO Plasmid (m) | sc-437372 | 20 µg | $397.00 |
Overview
Map3k20 encodes mixed-lineage kinase-like protein (MLTK), a serine/threonine MAP3K that links upstream stress and inflammatory cues to MAPK signaling cascades, including JNK and p38 pathways. Through phosphorylation of downstream MAP2Ks, MLTK contributes to regulation of transcriptional programs controlling apoptosis, cytoskeletal remodeling, and cellular adaptation to environmental stress. In mouse systems, Map3k20 activity has been associated with cell fate decisions in response to genotoxic and oxidative stress and can influence signaling nodes that intersect with innate immune and tissue homeostasis pathways. Dysregulated MAPK pathway wiring involving MAP3Ks like MLTK is broadly relevant to models of neurodegeneration, inflammatory pathology, and cancer-associated stress signaling, making Map3k20 a useful target for mechanistic dissection.
MLTK CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Map3k20 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Map3k20 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 Map3k20 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 MLTK protein expression.
This CRISPR knockout system enables efficient generation of Map3k20-deficient cell models for investigation of MLTK signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Map3k20 exon(s) critical for MLTK 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 Map3k20 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by MLTK CRISPR/Cas9 KO Plasmid (m) and MLTK CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Map3k20 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by MLTK HDR Plasmid (m) and MLTK HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Map3k20 homology arms to support homology-directed repair at defined Map3k20 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.