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.
Ubiquitin CRISPR/Cas9 KO Plasmid (m): sc-423574
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Ubiquitin 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 Ubiquitin 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: Ubiquitin Antibody (A-5): sc-166553
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Ubiquitin CRISPR/Cas9 KO Plasmid (m) | sc-423574 | 20 µg | $397.00 |
Overview
Ubb encodes ubiquitin, a highly conserved 76–amino acid modifier that is covalently attached to target proteins to regulate stability, localization, and signaling. Ubiquitination coordinates proteasome-dependent degradation, endocytosis, and DNA damage responses, and it interfaces with autophagy through ubiquitin-binding adaptors to control proteostasis. In mouse cells, ubiquitin-dependent signaling contributes broadly to cell-cycle progression, stress responses, and immune pathway regulation via ubiquitin chain topology and E3 ligase specificity. Dysregulated ubiquitin homeostasis and proteasome flux are commonly leveraged as mechanistic readouts in studies of neurodegeneration, inflammation, and cancer-relevant stress adaptation, making Ubb a key node for pathway interrogation.
Ubiquitin CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Ubb gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Ubb 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 Ubb 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 Ubiquitin protein expression.
This CRISPR knockout system enables efficient generation of Ubb-deficient cell models for investigation of Ubiquitin signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Ubb exon(s) critical for Ubiquitin 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 Ubb genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Ubiquitin CRISPR/Cas9 KO Plasmid (m) and Ubiquitin CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Ubb locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Ubiquitin HDR Plasmid (m) and Ubiquitin HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Ubb homology arms to support homology-directed repair at defined Ubb 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.