HLA-DQB1 CRISPR/Cas9 KO Plasmid (m): sc-420756

Overview

H2-Ab1 encodes the murine MHC class II beta chain functionally analogous to human HLA-DQB1 and forms a heterodimer that presents extracellular peptide antigens to CD4+ T cells. This antigen presentation axis supports thymic selection, peripheral T cell activation, and maintenance of immune tolerance through regulated trafficking and peptide loading in the endosomal pathway. H2-Ab1-dependent signaling influences cytokine programs and downstream adaptive immune responses, linking it to mechanisms that shape inflammation and autoimmunity. Dysregulated MHC class II expression or peptide presentation has been widely associated with altered susceptibility to immune-mediated pathology and variability in antigen-specific immune responses in mouse disease models.

HLA-DQB1 CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the H2-Ab1 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the H2-Ab1 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 H2-Ab1 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 HLA-DQB1 protein expression.

This CRISPR knockout system enables efficient generation of H2-Ab1-deficient cell models for investigation of HLA-DQB1 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.

Key Features

• sgRNAs targeting H2-Ab1 exon(s) critical for HLA-DQB1 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 H2-Ab1 genomic sites to improve knockout efficiency
• Compatible with delivery by transfection

Design Variants

CRISPRs +/- HDRs

• gRNAs encoded by HLA-DQB1 CRISPR/Cas9 KO Plasmid (m) and HLA-DQB1 CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the H2-Ab1 locus. One or both targeting designs may be available. See Related Products for availability.
• HDR donor constructs encoded by HLA-DQB1 HDR Plasmid (m) and HLA-DQB1 HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by H2-Ab1 homology arms to support homology-directed repair at defined H2-Ab1 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.