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.
Rag A CRISPR/Cas9 KO Plasmid (h): sc-411501
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Rag A CRISPR/Cas9 Knockout (KO) Plasmid (h) 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 Rag A 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: Rag A Antibody (D-1): sc-518209
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Rag A CRISPR/Cas9 KO Plasmid (h) | sc-411501 | 20 µg | $397.00 |
Overview
RRAGA encodes Rag A, a Ras-related small GTPase that forms heterodimers with RagC/D to couple intracellular amino acid availability to activation of mTORC1 at the lysosomal surface. Through regulated cycling between GDP- and GTP-bound states and interaction with the Ragulator complex and GATOR regulators, Rag A controls mTORC1 substrate phosphorylation, coordinating protein synthesis, autophagy suppression, and metabolic reprogramming. Dysregulated Rag GTPase signaling and aberrant mTORC1 activity are implicated in cancer biology and metabolic and neurodevelopmental disease contexts, making RRAGA a useful node for dissecting nutrient-sensing circuitry. Genetic perturbation of RRAGA enables mechanistic studies of lysosome-dependent signaling and downstream translational and autophagy pathways.
Rag A CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the RRAGA gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the RRAGA 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 RRAGA 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 Rag A protein expression.
This CRISPR knockout system enables efficient generation of RRAGA-deficient cell models for investigation of Rag A signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting RRAGA exon(s) critical for Rag A 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 RRAGA genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Rag A CRISPR/Cas9 KO Plasmid (h) and Rag A CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the RRAGA locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Rag A HDR Plasmid (h) and Rag A HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by RRAGA homology arms to support homology-directed repair at defined RRAGA 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.