Analysis Of Lymphocytes Development And Function In Gas41 Gene Knockout Mice

Research purpose:Gas41 gene deficiency causes the decrease in innate lymphoid cells（ILCs）,which suggests that it is required for the differentiation progress in common progenitor cells towards ILCs.However,how it impacts on B cell growth and differentiation is still unknown.To find out the function of Gas41 on lymphocytes development,hematopoietic stem cells and B cells’function,we constructed the Gas41gene knockout mouse model by using CRISPR/Cas9 gene editing technology.To study if Gas41 gene influences the formation of germinal center and the function of B cells,we used SRBC and NP₂₀-CGG to immunostimulate mice.Our study would provide a basis for the further study of Gas41 in immune regulation.Research method:we constructed the Gas41 gene knockout mouse model by using CRISPR/Cas9 gene editing technology.Both our experiment results and others find that knocking out the Gas41 gene allele would cause early embryos lethality in mice,so we use Gas41^+/-mice for further experiments.We use flow cytometry to find out the function of Gas41 on lymphocytes development,hematopoietic stem cells and B cells’function in Gas41^+/+and Gas41^+/-mice.To study if Gas41 gene influences the formation of germinal center and the function of B cells,we used SRBC and NP₂₀-CGG to immunostimulate Gas41^+/+and Gas41^+/-mice.Research results:In comparison with Gas41^+/+wild type mice,the proportion and the cell numbers of transitional T1 B cells in the spleen of Gas41^+/-mice are reduced,while the proportion of CD4⁺T,CD8⁺T cells and those cell numbers are increased.In addition,Ig G1 in serum is increased in Gas41^+/-mice.We found that anti-NP specific Ig G1 in serum is also increased in Gas41^+/-mice following NP₂₀-CGG immunolization while is not affected by SRBC stimulation.Conclusion:Down expression of Gas41 affects the developmental progress of T1 B cells,CD4⁺T and CD8⁺T cells in the spleen of Gas41^+/-mice.Down expression of Gas41 affects Ig G1 in basic serum,and Gas41^+/-mice would produce more anti-NP specific Ig G1 following NP₂₀-CGG immunolization.