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.
p130 CRISPR/Cas9 KO Plasmid (m): sc-422615
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- p130 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 p130 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: p130 Antibody (A-10): sc-374521
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
p130 CRISPR/Cas9 KO Plasmid (m) | sc-422615 | 20 µg | $397.00 |
Overview
Mouse Rbl2 encodes the retinoblastoma-like protein p130 (RBL2), a pocket protein that partners with E2F4/5 and DREAM complexes to repress transcription and enforce cell-cycle exit. p130 integrates signals from CDK-cyclin activity and phosphorylation-dependent remodeling of RB-family complexes to regulate the G0/G1 transition, quiescence, and differentiation programs. Through these pathways, Rbl2 influences checkpoint control, chromatin-associated repression, and lineage stability, making it relevant to studies of proliferation control and oncogenic deregulation. Altered RB-pathway function that perturbs p130-mediated E2F repression is frequently examined in models of cancer, senescence, and tissue homeostasis.
p130 CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Rbl2 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Rbl2 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 Rbl2 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 p130 protein expression.
This CRISPR knockout system enables efficient generation of Rbl2-deficient cell models for investigation of p130 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Rbl2 exon(s) critical for p130 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 Rbl2 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by p130 CRISPR/Cas9 KO Plasmid (m) and p130 CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Rbl2 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by p130 HDR Plasmid (m) and p130 HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Rbl2 homology arms to support homology-directed repair at defined Rbl2 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.