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.
RANKL CRISPR/Cas9 KO Plasmid (m): sc-423448
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- RANKL 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 RANKL 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: RANKL Antibody (12A668): sc-52950
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
RANKL CRISPR/Cas9 KO Plasmid (m) | sc-423448 | 20 µg | $397.00 |
Overview
Mouse Tnfsf11 encodes RANKL, a TNF superfamily ligand expressed by osteoblast-lineage stromal cells and activated immune cells that signals through its receptor RANK (TNFRSF11A) to control osteoclast differentiation, activation, and survival. RANKL–RANK engagement drives NF-κB, MAPK, and NFATc1-dependent transcriptional programs central to bone remodeling, lymph node organogenesis, and dendritic cell–T cell crosstalk. Physiologically, RANKL activity is counterbalanced by osteoprotegerin (OPG/TNFRSF11B) and contributes to coupling of immune activation with skeletal turnover. Dysregulated RANKL signaling is implicated in inflammatory bone erosion, osteoporosis-related bone loss, and osteolytic tumor–bone interactions, making it a key target for mechanistic studies of osteoimmunology.
RANKL CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Tnfsf11 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Tnfsf11 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 Tnfsf11 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 RANKL protein expression.
This CRISPR knockout system enables efficient generation of Tnfsf11-deficient cell models for investigation of RANKL signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Tnfsf11 exon(s) critical for RANKL 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 Tnfsf11 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by RANKL CRISPR/Cas9 KO Plasmid (m) and RANKL CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Tnfsf11 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by RANKL HDR Plasmid (m) and RANKL HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Tnfsf11 homology arms to support homology-directed repair at defined Tnfsf11 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.