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.
brachyury CRISPR/Cas9 KO Plasmid (m): sc-423247
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- brachyury 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 brachyury 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: brachyury Antibody (D-10): sc-166962
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
brachyury CRISPR/Cas9 KO Plasmid (m) | sc-423247 | 20 µg | $397.00 |
Overview
Mouse T encodes brachyury, a T-box transcription factor essential for mesoderm formation, primitive streak progression, and posterior body axis development during embryogenesis. Brachyury regulates lineage specification programs and coordinates transcriptional networks that control cell fate decisions, tissue patterning, and morphogenetic movements, integrating with signaling inputs such as WNT, FGF, and TGF-β/SMAD pathways. Dysregulated brachyury activity has been linked to aberrant mesenchymal differentiation and epithelial–mesenchymal transition-associated gene expression, supporting its relevance to developmental defects and disease-associated transcriptional reprogramming.
brachyury CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the T gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the T 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 T 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 brachyury protein expression.
This CRISPR knockout system enables efficient generation of T-deficient cell models for investigation of brachyury signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting T exon(s) critical for brachyury 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 T genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by brachyury CRISPR/Cas9 KO Plasmid (m) and brachyury CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the T locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by brachyury HDR Plasmid (m) and brachyury HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by T homology arms to support homology-directed repair at defined T 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.