HLA-DOβ CRISPR/Cas9 KO Plasmid (h): sc-406402
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- HLA-DOβ CRISPR/Cas9 Knockout (KO) Plasmid (h) 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 HLA-DOβ 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: HLA-DOβ Antibody (DOB.L1): sc-69739
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
HLA-DOβ CRISPR/Cas9 KO Plasmid (h) | sc-406402 | 20 µg | $397.00 |
Overview
HLA-DOB encodes the HLA-DOβ subunit, which pairs with HLA-DOα to form the HLA-DO heterodimer expressed primarily in antigen-presenting cells. Within late endosomes and lysosomes, HLA-DO modulates HLA-DM activity to fine-tune peptide loading onto MHC class II molecules, shaping the repertoire of peptides displayed to CD4+ T cells. This regulation links HLA-DOβ to antigen processing and presentation, adaptive immune tolerance, and immune responses to infection. Altered MHC class II peptide editing and HLA region variation are relevant to studies of autoimmunity, inflammatory disease mechanisms, and tumor–immune interactions where antigen presentation quality influences immune recognition.
HLA-DOβ CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the HLA-DOB gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the HLA-DOB 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 HLA-DOB 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-DOβ protein expression.
This CRISPR knockout system enables efficient generation of HLA-DOB-deficient cell models for investigation of HLA-DOβ signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting HLA-DOB exon(s) critical for HLA-DOβ 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 HLA-DOB genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by HLA-DOβ CRISPR/Cas9 KO Plasmid (h) and HLA-DOβ CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the HLA-DOB locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by HLA-DOβ HDR Plasmid (h) and HLA-DOβ HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by HLA-DOB homology arms to support homology-directed repair at defined HLA-DOB 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.