Creative BioMart provides MHC knockout cell line construction services based on CRISPR/Cas9 system gene-editing technology, which can achieve targeted editing of MHC molecular regions and obtain MHC molecular knockout cell lines. Our molecular biology team provides comprehensive gene knockout strategy design and subsequent cell line evaluation services to help you and your research get off to a fast start.

Methods of MHC Knockout

T lymphocytes of the adaptive immune system recognize short peptides derived from endogenous cellular proteins or exogenous antigens presented by MHC I and MHC II molecules on the cell surface. MHC class I (MHC I) molecules are expressed by any nucleated cell, expression of MHC class II molecules (MHC II) is mainly restricted to antigen-presenting cells such as macrophages, B cells or dendritic cells. The pool of MHC I-restricted peptides is permanently scanned by CD8⁺ cytotoxic T cells (CTLs) and, when activated upon recognition of exogenous peptides, will ultimately eradicate epitope-presenting target cells. In contrast, natural killer (NK) cells preferentially attack target cells that do not have MHC I surface expression but display activated NK cell receptors on their surface. Since the formation of stable MHC I/peptide complexes are heavily dependent on binding to β2m (light chain), defective β2m expression will abolish stable MHC I cell surface expression. Similar to MHC I-restricted epitope presentation, peptides presented by MHC II molecules are screened by CD4⁺ to recognize T cells activated after exogenous peptides. Since functional MHC II molecules are composed of α:β heterodimers, inactivation or deletion of either peptide chain (α or β) prevents their cell surface expression.

Construction of MHC Knockout Cell Lines

Our cell line construction services include transient and stable strains, which can be selected according to customer needs. MHC knockout affects the process of antigen presentation and is helpful in studying related immune dysfunction. The service constructs MHC knockout cell lines through CRISPR-Cas9-mediated multiplex gene editing for subsequent analytical testing. Our molecular biology team can provide services from gene editing protocol design to cell line testing. Customers can specify targeted knockout of heavy or light chain-related regions and result in underexpression of their surface MHC molecules. The CRISPR/Cas9 system is an efficient and direct strategy to generate stable MHC knockout cell lines such as tumor cell lines. The resulting MHCI/II-deficient cell lines can be used as parental lines for the establishment of MHC-compatible, transplantable tumor models in HLA transgenic mouse lines lacking expression of endogenous MHC molecules. MHC knockout cell lines represent a useful tool to study tumor-reactive T cell responses. Additionally, we offer a range of cell evaluation services to help you validate your modified cell lines for production.

Fig.1 MHC Knockout Cell Line Construction Services.

Creative BioMart has accumulated a lot of testing experience and can provide high-quality gene-editing services to achieve the purpose of targeted gene editing. We will help customers to achieve the knockout of MHC molecules. The knockout scheme based on the CRISPR/Cas9 system is classic and efficient, helping customers to quickly advance their research. If you are interested in our services, please contact us for more detailed information.

Reference

• Liu, Xiaojuan, et al. "CRISPR-Cas9-mediated multiplex gene editing in CAR-T cells." Cell research 27.1 (2017): 154-157.