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.
TIM CRISPR/Cas9 KO Plasmid (m): sc-423480
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- TIM 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 TIM 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: TIM Antibody (H-11): sc-166785
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
TIM CRISPR/Cas9 KO Plasmid (m) | sc-423480 | 20 µg | $397.00 |
Overview
Mouse Tpi1 encodes triosephosphate isomerase (TIM), a core glycolytic enzyme that catalyzes the reversible interconversion of dihydroxyacetone phosphate and glyceraldehyde-3-phosphate. By maintaining efficient flux through glycolysis, TIM supports cellular ATP production, redox balance, and metabolic coupling to pathways such as the pentose phosphate pathway and anaplerotic inputs into the TCA cycle. Tpi1 activity is therefore integral to bioenergetic homeostasis in proliferative and differentiated tissues, and its dysregulation is relevant to studies of metabolic stress responses, hypoxia adaptation, and glycolysis-dependent phenotypes. Altered glycolytic control involving Tpi1 is also commonly examined in models of neurological dysfunction, hemolytic pathology, and cancer-associated metabolic reprogramming.
TIM CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Tpi1 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Tpi1 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 Tpi1 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 TIM protein expression.
This CRISPR knockout system enables efficient generation of Tpi1-deficient cell models for investigation of TIM signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Tpi1 exon(s) critical for TIM 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 Tpi1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by TIM CRISPR/Cas9 KO Plasmid (m) and TIM CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Tpi1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by TIM HDR Plasmid (m) and TIM HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Tpi1 homology arms to support homology-directed repair at defined Tpi1 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.