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.
Smad6 CRISPR/Cas9 KO Plasmid (m): sc-421529
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Smad6 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 Smad6 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: Smad6 Antibody (D-4): sc-25321
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Smad6 CRISPR/Cas9 KO Plasmid (m) | sc-421529 | 20 µg | $397.00 |
Overview
SMAD6 is an inhibitory SMAD that attenuates signaling downstream of BMP and, in some contexts, TGF-β receptors by interfering with receptor-regulated SMAD phosphorylation and complex formation. In mouse cells, Smad6 helps tune BMP-driven transcriptional programs that govern developmental patterning, osteogenic and chondrogenic differentiation, and vascular homeostasis. By modulating canonical SMAD signaling, SMAD6 contributes to control of extracellular matrix remodeling, cell fate decisions, and feedback regulation of morphogen pathways. Dysregulated Smad6 activity has been linked to abnormal BMP pathway output and phenotypes relevant to congenital cardiovascular and skeletal biology, making it a useful node for mechanistic studies of pathway restraint.
Smad6 CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Smad6 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Smad6 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 Smad6 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 Smad6 protein expression.
This CRISPR knockout system enables efficient generation of Smad6-deficient cell models for investigation of Smad6 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Smad6 exon(s) critical for Smad6 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 Smad6 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Smad6 CRISPR/Cas9 KO Plasmid (m) and Smad6 CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Smad6 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Smad6 HDR Plasmid (m) and Smad6 HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Smad6 homology arms to support homology-directed repair at defined Smad6 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.