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.
CAD CRISPR/Cas9 KO Plasmid (h): sc-403028
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- CAD CRISPR/Cas9 Knockout (KO) Plasmid (h) 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 CAD 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: CAD Antibody (F-11): sc-374067
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
CAD CRISPR/Cas9 KO Plasmid (h) | sc-403028 | 20 µg | $397.00 |
Overview
DFFB encodes caspase-activated DNase (CAD), the endonuclease responsible for internucleosomal DNA fragmentation during apoptosis. In viable cells, CAD is maintained inactive through binding to its inhibitor and chaperone ICAD (DFFA), and becomes activated following caspase-3–dependent ICAD cleavage, enabling chromatin degradation and apoptotic nuclear dismantling. This nuclease activity links the intrinsic and extrinsic apoptosis pathways to genome integrity control by ensuring orderly DNA disposal during programmed cell death. Dysregulation of CAD/ICAD function can perturb apoptotic execution and has been investigated in contexts of oncogenic transformation, immune cell homeostasis, and tissue degeneration where altered cell death programs are implicated.
CAD CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the DFFB gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the DFFB 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 DFFB 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 CAD protein expression.
This CRISPR knockout system enables efficient generation of DFFB-deficient cell models for investigation of CAD signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting DFFB exon(s) critical for CAD 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 DFFB genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by CAD CRISPR/Cas9 KO Plasmid (h) and CAD CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the DFFB locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by CAD HDR Plasmid (h) and CAD HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by DFFB homology arms to support homology-directed repair at defined DFFB 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.