CD3-ζ/η CRISPR/Cas9 KO Plasmid (m): sc-419556

Overview

Cd247 encodes the CD3ζ/η signaling subunit of the T cell receptor (TCR) complex, an immunoreceptor tyrosine-based activation motif (ITAM)-containing adaptor that couples antigen recognition to intracellular phosphorylation cascades. Upon TCR engagement, CD3ζ/η ITAMs recruit and activate kinases such as LCK and ZAP70, promoting LAT/SLP-76 signalosome assembly and downstream MAPK, NF-κB, and NFAT pathways that control T cell activation, cytokine production, and survival. Altered CD247 expression or signaling contributes to impaired T cell responsiveness and immune dysregulation, and has been linked to inflammatory and autoimmune phenotypes as well as tumor immune evasion in the microenvironment. In mice, Cd247 perturbation is widely used to dissect thymic selection, peripheral tolerance, and effector versus exhausted T cell states.

CD3-ζ/η CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Cd247 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Cd247 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 Cd247 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 CD3-ζ/η protein expression.

This CRISPR knockout system enables efficient generation of Cd247-deficient cell models for investigation of CD3-ζ/η signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.

Key Features

• sgRNAs targeting Cd247 exon(s) critical for CD3-ζ/η 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 Cd247 genomic sites to improve knockout efficiency
• Compatible with delivery by transfection

Design Variants

CRISPRs +/- HDRs

• gRNAs encoded by CD3-ζ/η CRISPR/Cas9 KO Plasmid (m) and CD3-ζ/η CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Cd247 locus. One or both targeting designs may be available. See Related Products for availability.
• HDR donor constructs encoded by CD3-ζ/η HDR Plasmid (m) and CD3-ζ/η HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Cd247 homology arms to support homology-directed repair at defined Cd247 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.