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.
dUTPase CRISPR/Cas9 KO Plasmid (h): sc-405843
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- dUTPase 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 dUTPase 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: dUTPase Antibody (H-9): sc-166856
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
dUTPase CRISPR/Cas9 KO Plasmid (h) | sc-405843 | 20 µg | $397.00 |
Overview
DUT encodes human dUTPase, a key nucleotide metabolism enzyme that hydrolyzes dUTP to dUMP and pyrophosphate, thereby maintaining low intracellular dUTP pools and supplying substrate for thymidylate biosynthesis. By limiting uracil incorporation into genomic DNA, dUTPase supports replication fidelity and genome stability and interfaces functionally with base excision repair pathways that remove uracil from DNA. Altered DUT activity can modulate replication stress responses and DNA damage signaling, processes frequently perturbed in proliferative disorders and in cells challenged by genotoxic stress. As a central node in pyrimidine homeostasis, DUT is widely studied in the context of cell cycle progression, mitochondrial and nuclear DNA maintenance, and nucleotide imbalance–driven mutagenesis.
dUTPase CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the DUT gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the DUT 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 DUT 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 dUTPase protein expression.
This CRISPR knockout system enables efficient generation of DUT-deficient cell models for investigation of dUTPase signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting DUT exon(s) critical for dUTPase 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 DUT genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by dUTPase CRISPR/Cas9 KO Plasmid (h) and dUTPase CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the DUT locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by dUTPase HDR Plasmid (h) and dUTPase HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by DUT homology arms to support homology-directed repair at defined DUT 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.