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.
TS CRISPR/Cas9 KO Plasmid (m): sc-423565
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- TS 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 TS 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: TS Antibody (C-5): sc-390945
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
TS CRISPR/Cas9 KO Plasmid (m) | sc-423565 | 20 µg | $397.00 |
Overview
Tyms encodes thymidylate synthase (TS), a key folate-dependent enzyme that catalyzes the conversion of dUMP to dTMP, maintaining thymidine nucleotide pools required for DNA replication and repair. TS activity links one-carbon metabolism to S-phase progression, genome stability, and mitochondrial and nuclear DNA synthesis through the de novo pyrimidine biosynthesis network. In mouse systems, Tyms regulation and TS abundance are commonly studied in proliferative tissues and rapidly cycling cell models where nucleotide imbalance can trigger replication stress and DNA damage signaling. Dysregulated TS-associated metabolic flux is relevant to mechanisms of altered cell cycle control and genomic instability that are frequently interrogated in cancer biology and folate pathway research.
TS CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Tyms gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Tyms 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 Tyms 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 TS protein expression.
This CRISPR knockout system enables efficient generation of Tyms-deficient cell models for investigation of TS signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Tyms exon(s) critical for TS 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 Tyms genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by TS CRISPR/Cas9 KO Plasmid (m) and TS CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Tyms locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by TS HDR Plasmid (m) and TS HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Tyms homology arms to support homology-directed repair at defined Tyms 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.