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.
Ral A CRISPR/Cas9 KO Plasmid (h): sc-403983
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Ral 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 Ral 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: Ral A Antibody (F-5): sc-374462
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Ral A CRISPR/Cas9 KO Plasmid (h) | sc-403983 | 20 µg | $397.00 |
Overview
RALA encodes RalA, a Ras-related small GTPase that cycles between GDP- and GTP-bound states to coordinate signal transduction downstream of Ras family inputs. Active RalA regulates vesicle trafficking, polarized exocytosis, endocytosis, and actin cytoskeleton remodeling through effectors such as the exocyst complex and RalBP1, linking membrane dynamics to cell migration and proliferation. RalA signaling interfaces with MAPK- and PI3K-associated networks and contributes to control of cytokinesis and mitochondrial homeostasis. Dysregulated RALA activity has been implicated in oncogenic signaling contexts and altered cell motility programs, making it relevant for mechanistic studies of tumor biology and metastasis-associated pathways.
Ral A CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the RALA gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the RALA 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 RALA 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 Ral A protein expression.
This CRISPR knockout system enables efficient generation of RALA-deficient cell models for investigation of Ral A signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting RALA exon(s) critical for Ral 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 RALA genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Ral A CRISPR/Cas9 KO Plasmid (h) and Ral A CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the RALA locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Ral A HDR Plasmid (h) and Ral A HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by RALA homology arms to support homology-directed repair at defined RALA 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.