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| Item Type: | Article |
|---|---|
| Title: | Efficient CRISPR/Cas9-mediated mutagenesis in primary human B cells for identifying plasma cell regulators |
| Creators Name: | Le, T.A., Chu, V.T., Lino, A.C., Schrezenmeier, E., Kressler, C., Hamo, D., Rajewsky, K., Dörner, T. and Dang, V.D. |
| Abstract: | Human B lymphocytes are attractive targets for immunotherapies in autoantibody-mediated diseases. Gene editing technologies could provide a powerful tool to determine gene regulatory 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 approximate 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. Specifically, that IRF4, PRDM1 and XBP1 are expressed at different stages during plasma cell differentiation, IRF4, PRDM1 and XBP1-targeted cells fail to progress to the pre-PB, PC state and PC 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. |
| Keywords: | CRISPR/Cas9, Primary Human B Cells, B Cell Differentiation, Plasmablasts, Plasma Cells, Plasma Cell Regulators, Knockout, IRF4, PRDM1, XBP1 |
| Source: | Molecular Therapy - Nucleic Acids |
| ISSN: | 2162-2531 |
| Publisher: | Nature Publishing Group |
| Volume: | 30 |
| Page Range: | 621-632 |
| Date: | 13 December 2022 |
| Official Publication: | https://doi.org/10.1016/j.omtn.2022.11.016 |
| PubMed: | View item in PubMed |
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