[Construction of β2m knockout mice]

Area of Science:

• Immunology
• Genetics
• Molecular Biology

Background:

• Beta-2-microglobin (β2m) is crucial for Major Histocompatibility Complex class I (MHC I) molecule function in antigen presentation.
• Tumors and pathogens can evade immune detection by downregulating MHC I expression.
• Development of precise animal models is vital for studying diseases and therapeutic strategies.

Purpose of the Study:

• To generate and validate a β2m knockout mouse model.
• To investigate the role of β2m in immune responses.
• To provide a tool for in vivo research on β2m function and related diseases.

Main Methods:

• CRISPR/Cas9 gene editing was employed for β2m knockout mouse generation via embryo microinjection.
• Genotyping was confirmed using Polymerase Chain Reaction (PCR).
• Quantitative PCR (qPCR) and flow cytometry were used for phenotyping and assessing gene/protein expression and immune cell populations.

Main Results:

• Successful deletion of the β2m coding region in knockout mice.
• Significant downregulation of β2m mRNA levels was observed.
• A marked reduction in CD8+ killer T cell populations was detected in various tissues.

Conclusions:

• A novel strain of β2m knockout mice has been successfully established.
• This model demonstrates impaired MHC I antigen presentation due to β2m deficiency.
• The generated mouse model is a valuable resource for future in vivo studies on β2m's role in immunity, cancer, and disease.