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.
RLF CRISPR/Cas9 KO Plasmid (h): sc-406295
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- RLF 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 RLF 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: RLF Antibody (D-7): sc-365373
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
RLF CRISPR/Cas9 KO Plasmid (h) | sc-406295 | 20 µg | $397.00 |
Overview
RGL2 (Ral guanine nucleotide dissociation stimulator-like 2) encodes a Ras family guanine nucleotide exchange factor that helps activate RalA/RalB small GTPases downstream of Ras signaling. Through modulation of Ral GTPase cycling, RGL2 influences MAPK- and PI3K-linked signaling outputs that govern cytoskeletal remodeling, vesicle trafficking, cell-cycle progression, and stress responses. Altered activity within Ras–Ral signaling has been implicated in oncogenic transformation and tumor cell invasion in multiple tissue contexts, making RGL2 a useful node for mechanistic studies of pathway wiring. In human cell models, perturbing RGL2 supports investigation of how Ras-effector selection shapes proliferative and migratory phenotypes and intersects with receptor tyrosine kinase signaling.
RLF CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the RGL2 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the RGL2 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 RGL2 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 RLF protein expression.
This CRISPR knockout system enables efficient generation of RGL2-deficient cell models for investigation of RLF signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting RGL2 exon(s) critical for RLF 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 RGL2 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by RLF CRISPR/Cas9 KO Plasmid (h) and RLF CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the RGL2 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by RLF HDR Plasmid (h) and RLF HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by RGL2 homology arms to support homology-directed repair at defined RGL2 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.