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.
Mad 1 CRISPR/Cas9 KO Plasmid (m): sc-421519
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Mad 1 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 Mad 1 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: Mad 1 Antibody (F-1): sc-8012
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Mad 1 CRISPR/Cas9 KO Plasmid (m) | sc-421519 | 20 µg | $397.00 |
Overview
Mxd1 (Mad1) encodes a basic helix–loop–helix leucine zipper transcriptional regulator that antagonizes MYC-driven gene expression by partnering with MAX and recruiting corepressor complexes. Through this MYC–MAX–MAD network, Mad1 contributes to transcriptional repression programs controlling cell-cycle progression, differentiation, and cellular metabolism. In mouse systems, altered Mxd1 activity is studied in contexts where MYC signaling is deregulated, including oncogenic transformation, hematopoietic phenotypes, and developmental patterning. Mad1-dependent regulation of chromatin and RNA polymerase II output makes it a useful node for dissecting proliferative versus differentiative transcriptional states.
Mad 1 CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Mxd1 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Mxd1 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 Mxd1 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 Mad 1 protein expression.
This CRISPR knockout system enables efficient generation of Mxd1-deficient cell models for investigation of Mad 1 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Mxd1 exon(s) critical for Mad 1 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 Mxd1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Mad 1 CRISPR/Cas9 KO Plasmid (m) and Mad 1 CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Mxd1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Mad 1 HDR Plasmid (m) and Mad 1 HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Mxd1 homology arms to support homology-directed repair at defined Mxd1 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.