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.
53BP1 CRISPR/Cas9 KO Plasmid (m2): sc-424212-KO-2
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- 53BP1 CRISPR/Cas9 Knockout (KO) Plasmid (m2) 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 53BP1 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: p-53BP1 Antibody (38.Ser 25): sc-135748
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
53BP1 CRISPR/Cas9 KO Plasmid (m2) | sc-424212-KO-2 | 20 µg | $397.00 |
Overview
Trp53bp1 encodes 53BP1, a chromatin-associated mediator of the DNA damage response that is rapidly recruited to double-strand breaks via recognition of histone marks and ubiquitin signaling. 53BP1 promotes non-homologous end joining and limits DNA end resection, thereby influencing pathway choice between NHEJ and homologous recombination during repair. It coordinates checkpoint signaling, genomic stability, and class switch recombination in B cells, and its dysfunction is linked to elevated chromosomal instability and altered responses to replication stress. In mouse models, perturbation of Trp53bp1 is widely used to study genome maintenance, tumor suppressor networks, and BRCA1-associated repair pathway balance.
53BP1 CRISPR/Cas9 KO Plasmid (m2) is a pool of plasmids designed for targeted disruption of the Trp53bp1 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Trp53bp1 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 Trp53bp1 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 53BP1 protein expression.
This CRISPR knockout system enables efficient generation of Trp53bp1-deficient cell models for investigation of 53BP1 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Trp53bp1 exon(s) critical for 53BP1 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 Trp53bp1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by 53BP1 CRISPR/Cas9 KO Plasmid (m) and 53BP1 CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Trp53bp1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by 53BP1 HDR Plasmid (m) and 53BP1 HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Trp53bp1 homology arms to support homology-directed repair at defined Trp53bp1 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.