期刊
MOLECULAR THERAPY-NUCLEIC ACIDS
卷 30, 期 -, 页码 621-632出版社
CELL PRESS
DOI: 10.1016/j.omtn.2022.11.016
关键词
-
资金
- German Research Foundation [Do491/8-1/2, TRR130/TP24, Do491/10-1, Do491/11-1, LI3540/1-1]
Gene editing using CRISPR-Cas9 technology in human B cells can help identify gene regulatory networks and therapeutic targets for autoimmune diseases. Targeting IRF4, PRDM1, and XBP1 genes severely impairs B cell survival and plasma cell differentiation. This method provides a new avenue for studying gene functions and identifying plasma cell regulators.
Human B lymphocytes are attractive targets for immunother-apies in autoantibody-mediated diseases. Gene editing technol-ogies could provide a powerful tool to determine gene regula-tory networks regulating B cell differentiation into plasma cells, and identify novel therapeutic targets for prevention and treatment of autoimmune disorders. Here, we describe a new approach that uses CRISPR-Cas9 technology to target genes in primary human B cells in vitro for identifying plasma cell regulators. We found that sgRNA and Cas9 components can be efficiently delivered into primary human B cells through RD114-pseudotyped retroviral vectors. Using this system, we achieved approximately 80% of gene knockout efficiency. We disrupted expression of a triad of transcription factors, IRF4, PRDM1, and XBP1, and showed that human B cell survival and plasma cell differentiation are severely impaired. Specif-ically, that IRF4, PRDM1, and XBP1 were expressed at different stages during plasma cell differentiation, IRF4, PRDM1, and XBP1-targeted B cells failed to progress to the pre-plasmablast, plasma cell state, and plasma cell survival, respectively. Our method opens a new avenue to study gene functions in primary human B cells and identify novel plasma cell regulators for therapeutic applications.
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