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.
Rheb CRISPR/Cas9 KO Plasmid (m): sc-422675
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Rheb 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 Rheb 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: Rheb Antibody (80-R): sc-130398
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Rheb CRISPR/Cas9 KO Plasmid (m) | sc-422675 | 20 µg | $397.00 |
Overview
Mouse Rheb encodes a Ras-related small GTPase that functions as a core activator of mTORC1 at lysosomal membranes, integrating growth factor signaling, amino acid sensing, and cellular energy status to regulate protein synthesis, autophagy, and cell growth. Through the TSC1/TSC2 complex, Rheb activity is tightly controlled by PI3K–AKT signaling and stress-responsive inputs, shaping anabolic metabolism and mitochondrial homeostasis. Dysregulated Rheb–mTORC1 signaling is implicated in neurodevelopmental phenotypes, tumor biology, and metabolic remodeling, making Rheb a widely used node for dissecting nutrient signaling and growth control. In murine systems, perturbing Rheb supports mechanistic studies of mTOR-dependent translation programs, lysosome-associated signaling, and stress adaptation.
Rheb CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Rheb gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Rheb 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 Rheb 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 Rheb protein expression.
This CRISPR knockout system enables efficient generation of Rheb-deficient cell models for investigation of Rheb signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Rheb exon(s) critical for Rheb 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 Rheb genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Rheb CRISPR/Cas9 KO Plasmid (m) and Rheb CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Rheb locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Rheb HDR Plasmid (m) and Rheb HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Rheb homology arms to support homology-directed repair at defined Rheb 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.