TPMT CRISPR/Cas9 KO Plasmid (m): sc-423483
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- TPMT 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 TPMT 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
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
TPMT CRISPR/Cas9 KO Plasmid (m) | sc-423483 | 20 µg | $397.00 |
Overview
Tpmt encodes thiopurine S-methyltransferase (TPMT), a cytosolic S-adenosyl-L-methionine–dependent methyltransferase that catalyzes S-methylation of thiopurine and related sulfur-containing substrates. In mouse cells, TPMT activity contributes to xenobiotic and nucleotide analog metabolism, shaping intracellular thioguanine nucleotide pools and downstream impacts on DNA replication stress and damage signaling. Variation in TPMT function is widely used as a model for studying determinants of thiopurine sensitivity and hematopoietic toxicity mechanisms without implying clinical application. Tpmt is therefore relevant to pathways linking drug biotransformation, redox balance, and genome integrity in proliferative tissues.
TPMT CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Tpmt gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Tpmt 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 Tpmt 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 TPMT protein expression.
This CRISPR knockout system enables efficient generation of Tpmt-deficient cell models for investigation of TPMT signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Tpmt exon(s) critical for TPMT 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 Tpmt genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by TPMT CRISPR/Cas9 KO Plasmid (m) and TPMT CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Tpmt locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by TPMT HDR Plasmid (m) and TPMT HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Tpmt homology arms to support homology-directed repair at defined Tpmt 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.