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.
UBE1 CRISPR/Cas9 KO Plasmid (h): sc-402844
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- UBE1 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 UBE1 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: UBE1 Antibody (2G2): sc-53555
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
UBE1 CRISPR/Cas9 KO Plasmid (h) | sc-402844 | 20 µg | $397.00 |
Overview
UBA1 encodes ubiquitin-like modifier activating enzyme 1 (UBE1), the E1 enzyme that initiates ATP-dependent ubiquitin activation and transfers ubiquitin to E2 conjugating enzymes to drive ubiquitin–proteasome system function. Through this upstream control point, UBE1 influences global protein turnover, proteostasis, and stress responses, impacting processes such as cell-cycle progression, DNA damage signaling, and regulation of innate immune pathways. Perturbation of UBA1 activity can remodel ubiquitin-dependent signaling networks and alter stability of key regulatory proteins, making it relevant to studies of neurodegeneration, cancer-associated proteostasis dependencies, and X-linked disorders including spinal muscular atrophy. Because UBE1 sits at the apex of ubiquitination, genetic disruption provides a direct approach to dissect ubiquitin pathway contributions to cellular homeostasis.
UBE1 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the UBA1 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the UBA1 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 UBA1 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 UBE1 protein expression.
This CRISPR knockout system enables efficient generation of UBA1-deficient cell models for investigation of UBE1 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting UBA1 exon(s) critical for UBE1 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 UBA1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by UBE1 CRISPR/Cas9 KO Plasmid (h) and UBE1 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the UBA1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by UBE1 HDR Plasmid (h) and UBE1 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by UBA1 homology arms to support homology-directed repair at defined UBA1 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.