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.
GβL CRISPR/Cas9 KO Plasmid (m): sc-425272
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GβL 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 GβL 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: GβL Antibody (E-9): sc-514982
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
GβL CRISPR/Cas9 KO Plasmid (m) | sc-425272 | 20 µg | $397.00 |
Overview
Mlst8 encodes the GβL (mLST8) protein, a core component of both mTORC1 and mTORC2 complexes that stabilizes the mTOR kinase and supports complex assembly and signaling output. Through these complexes, GβL influences nutrient and growth factor sensing, translational control, autophagy, and cytoskeletal regulation, integrating upstream PI3K–AKT and related pathways. In mouse systems, perturbation of Mlst8 is used to interrogate how mTOR signaling coordinates cell growth, metabolism, and stress responses across tissues and developmental contexts. Dysregulated mTOR pathway activity is widely implicated in proliferative and metabolic disease biology, making Mlst8 a useful node for mechanistic studies of pathway dependency and signaling network rewiring.
GβL CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Mlst8 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Mlst8 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 Mlst8 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 GβL protein expression.
This CRISPR knockout system enables efficient generation of Mlst8-deficient cell models for investigation of GβL signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Mlst8 exon(s) critical for GβL 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 Mlst8 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by GβL CRISPR/Cas9 KO Plasmid (m) and GβL CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Mlst8 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by GβL HDR Plasmid (m) and GβL HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Mlst8 homology arms to support homology-directed repair at defined Mlst8 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.