TIM-3 CRISPR/Cas9 KO Plasmid (m): sc-431363
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- TIM-3 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-3 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 | |
TIM-3 CRISPR/Cas9 KO Plasmid (m) | sc-431363 | 20 µg | $397.00 |
Overview
Havcr2 encodes the immunoregulatory receptor TIM-3, a type I membrane protein expressed on multiple immune lineages including activated T cells, regulatory T cells, dendritic cells, and innate immune subsets. TIM-3 integrates signals that shape T cell activation thresholds, tolerance, and effector differentiation, influencing cytokine production and immune synapse function in the context of antigen stimulation. Through interactions with ligands such as galectin-9 and phosphatidylserine, TIM-3 contributes to pathways associated with T cell dysfunction and chronic antigen exposure, aligning it with immune exhaustion programs and inflammatory circuit modulation. In mouse models, Havcr2/TIM-3 biology is widely interrogated in settings of autoimmunity, chronic infection, and tumor immunology to understand immune homeostasis and tissue-specific inflammation.
TIM-3 CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Havcr2 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Havcr2 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 Havcr2 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-3 protein expression.
This CRISPR knockout system enables efficient generation of Havcr2-deficient cell models for investigation of TIM-3 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Havcr2 exon(s) critical for TIM-3 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 Havcr2 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by TIM-3 CRISPR/Cas9 KO Plasmid (m) and TIM-3 CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Havcr2 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by TIM-3 HDR Plasmid (m) and TIM-3 HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Havcr2 homology arms to support homology-directed repair at defined Havcr2 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.