Efficient CRISPR-Cas9-mediated mutagenesis in primary human B cells for identifying plasma cell regulators

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 a...
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Autor*in:	Tuan Anh Le [verfasserIn] Van Trung Chu [verfasserIn] Andreia C. Lino [verfasserIn] Eva Schrezenmeier [verfasserIn] Christopher Kressler [verfasserIn] Dania Hamo [verfasserIn] Klaus Rajewsky [verfasserIn] Thomas Dörner [verfasserIn] Van Duc Dang [verfasserIn]

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2022

Schlagwörter:	MT: RNA/DNA Editing CRISPR-Cas9 primary human B cells B cell differentiation plasmablasts plasma cells

Übergeordnetes Werk:	In: Molecular Therapy: Nucleic Acids - Elsevier, 2013, 30(2022), Seite 621-632
Übergeordnetes Werk:	volume:30 ; year:2022 ; pages:621-632

Links:	Link aufrufen Link aufrufen Link aufrufen Journal toc

DOI / URN:	10.1016/j.omtn.2022.11.016

Katalog-ID:	DOAJ083776613

Internformat


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520			\|a 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 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. Specifically, 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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allfields	10.1016/j.omtn.2022.11.016 doi (DE-627)DOAJ083776613 (DE-599)DOAJ174f4a9188c14eeabc505f36d39a9a14 DE-627 ger DE-627 rakwb eng RM1-950 Tuan Anh Le verfasserin aut Efficient CRISPR-Cas9-mediated mutagenesis in primary human B cells for identifying plasma cell regulators 2022 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier 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 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. Specifically, 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. MT: RNA/DNA Editing CRISPR-Cas9 primary human B cells B cell differentiation plasmablasts plasma cells Therapeutics. Pharmacology Van Trung Chu verfasserin aut Andreia C. Lino verfasserin aut Eva Schrezenmeier verfasserin aut Christopher Kressler verfasserin aut Dania Hamo verfasserin aut Klaus Rajewsky verfasserin aut Thomas Dörner verfasserin aut Van Duc Dang verfasserin aut In Molecular Therapy: Nucleic Acids Elsevier, 2013 30(2022), Seite 621-632 (DE-627)718632702 (DE-600)2662631-7 21622531 nnns volume:30 year:2022 pages:621-632 https://doi.org/10.1016/j.omtn.2022.11.016 kostenfrei https://doaj.org/article/174f4a9188c14eeabc505f36d39a9a14 kostenfrei http://www.sciencedirect.com/science/article/pii/S2162253122003080 kostenfrei https://doaj.org/toc/2162-2531 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2007 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2026 GBV_ILN_2027 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2068 GBV_ILN_2088 GBV_ILN_2110 GBV_ILN_2112 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2153 GBV_ILN_2190 GBV_ILN_2336 GBV_ILN_2470 GBV_ILN_2507 GBV_ILN_4012 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4326 GBV_ILN_4333 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4393 GBV_ILN_4700 AR 30 2022 621-632
spelling	10.1016/j.omtn.2022.11.016 doi (DE-627)DOAJ083776613 (DE-599)DOAJ174f4a9188c14eeabc505f36d39a9a14 DE-627 ger DE-627 rakwb eng RM1-950 Tuan Anh Le verfasserin aut Efficient CRISPR-Cas9-mediated mutagenesis in primary human B cells for identifying plasma cell regulators 2022 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier 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 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. Specifically, 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. MT: RNA/DNA Editing CRISPR-Cas9 primary human B cells B cell differentiation plasmablasts plasma cells Therapeutics. Pharmacology Van Trung Chu verfasserin aut Andreia C. Lino verfasserin aut Eva Schrezenmeier verfasserin aut Christopher Kressler verfasserin aut Dania Hamo verfasserin aut Klaus Rajewsky verfasserin aut Thomas Dörner verfasserin aut Van Duc Dang verfasserin aut In Molecular Therapy: Nucleic Acids Elsevier, 2013 30(2022), Seite 621-632 (DE-627)718632702 (DE-600)2662631-7 21622531 nnns volume:30 year:2022 pages:621-632 https://doi.org/10.1016/j.omtn.2022.11.016 kostenfrei https://doaj.org/article/174f4a9188c14eeabc505f36d39a9a14 kostenfrei http://www.sciencedirect.com/science/article/pii/S2162253122003080 kostenfrei https://doaj.org/toc/2162-2531 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2007 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2026 GBV_ILN_2027 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2068 GBV_ILN_2088 GBV_ILN_2110 GBV_ILN_2112 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2153 GBV_ILN_2190 GBV_ILN_2336 GBV_ILN_2470 GBV_ILN_2507 GBV_ILN_4012 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4326 GBV_ILN_4333 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4393 GBV_ILN_4700 AR 30 2022 621-632
allfields_unstemmed	10.1016/j.omtn.2022.11.016 doi (DE-627)DOAJ083776613 (DE-599)DOAJ174f4a9188c14eeabc505f36d39a9a14 DE-627 ger DE-627 rakwb eng RM1-950 Tuan Anh Le verfasserin aut Efficient CRISPR-Cas9-mediated mutagenesis in primary human B cells for identifying plasma cell regulators 2022 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier 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 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. Specifically, 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. MT: RNA/DNA Editing CRISPR-Cas9 primary human B cells B cell differentiation plasmablasts plasma cells Therapeutics. Pharmacology Van Trung Chu verfasserin aut Andreia C. Lino verfasserin aut Eva Schrezenmeier verfasserin aut Christopher Kressler verfasserin aut Dania Hamo verfasserin aut Klaus Rajewsky verfasserin aut Thomas Dörner verfasserin aut Van Duc Dang verfasserin aut In Molecular Therapy: Nucleic Acids Elsevier, 2013 30(2022), Seite 621-632 (DE-627)718632702 (DE-600)2662631-7 21622531 nnns volume:30 year:2022 pages:621-632 https://doi.org/10.1016/j.omtn.2022.11.016 kostenfrei https://doaj.org/article/174f4a9188c14eeabc505f36d39a9a14 kostenfrei http://www.sciencedirect.com/science/article/pii/S2162253122003080 kostenfrei https://doaj.org/toc/2162-2531 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2007 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2026 GBV_ILN_2027 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2068 GBV_ILN_2088 GBV_ILN_2110 GBV_ILN_2112 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2153 GBV_ILN_2190 GBV_ILN_2336 GBV_ILN_2470 GBV_ILN_2507 GBV_ILN_4012 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4326 GBV_ILN_4333 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4393 GBV_ILN_4700 AR 30 2022 621-632
allfieldsGer	10.1016/j.omtn.2022.11.016 doi (DE-627)DOAJ083776613 (DE-599)DOAJ174f4a9188c14eeabc505f36d39a9a14 DE-627 ger DE-627 rakwb eng RM1-950 Tuan Anh Le verfasserin aut Efficient CRISPR-Cas9-mediated mutagenesis in primary human B cells for identifying plasma cell regulators 2022 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier 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 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. Specifically, 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. MT: RNA/DNA Editing CRISPR-Cas9 primary human B cells B cell differentiation plasmablasts plasma cells Therapeutics. Pharmacology Van Trung Chu verfasserin aut Andreia C. Lino verfasserin aut Eva Schrezenmeier verfasserin aut Christopher Kressler verfasserin aut Dania Hamo verfasserin aut Klaus Rajewsky verfasserin aut Thomas Dörner verfasserin aut Van Duc Dang verfasserin aut In Molecular Therapy: Nucleic Acids Elsevier, 2013 30(2022), Seite 621-632 (DE-627)718632702 (DE-600)2662631-7 21622531 nnns volume:30 year:2022 pages:621-632 https://doi.org/10.1016/j.omtn.2022.11.016 kostenfrei https://doaj.org/article/174f4a9188c14eeabc505f36d39a9a14 kostenfrei http://www.sciencedirect.com/science/article/pii/S2162253122003080 kostenfrei https://doaj.org/toc/2162-2531 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2007 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2026 GBV_ILN_2027 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2068 GBV_ILN_2088 GBV_ILN_2110 GBV_ILN_2112 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2153 GBV_ILN_2190 GBV_ILN_2336 GBV_ILN_2470 GBV_ILN_2507 GBV_ILN_4012 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4326 GBV_ILN_4333 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4393 GBV_ILN_4700 AR 30 2022 621-632
allfieldsSound	10.1016/j.omtn.2022.11.016 doi (DE-627)DOAJ083776613 (DE-599)DOAJ174f4a9188c14eeabc505f36d39a9a14 DE-627 ger DE-627 rakwb eng RM1-950 Tuan Anh Le verfasserin aut Efficient CRISPR-Cas9-mediated mutagenesis in primary human B cells for identifying plasma cell regulators 2022 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier 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 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. Specifically, 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. MT: RNA/DNA Editing CRISPR-Cas9 primary human B cells B cell differentiation plasmablasts plasma cells Therapeutics. Pharmacology Van Trung Chu verfasserin aut Andreia C. Lino verfasserin aut Eva Schrezenmeier verfasserin aut Christopher Kressler verfasserin aut Dania Hamo verfasserin aut Klaus Rajewsky verfasserin aut Thomas Dörner verfasserin aut Van Duc Dang verfasserin aut In Molecular Therapy: Nucleic Acids Elsevier, 2013 30(2022), Seite 621-632 (DE-627)718632702 (DE-600)2662631-7 21622531 nnns volume:30 year:2022 pages:621-632 https://doi.org/10.1016/j.omtn.2022.11.016 kostenfrei https://doaj.org/article/174f4a9188c14eeabc505f36d39a9a14 kostenfrei http://www.sciencedirect.com/science/article/pii/S2162253122003080 kostenfrei https://doaj.org/toc/2162-2531 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2007 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2026 GBV_ILN_2027 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2068 GBV_ILN_2088 GBV_ILN_2110 GBV_ILN_2112 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2153 GBV_ILN_2190 GBV_ILN_2336 GBV_ILN_2470 GBV_ILN_2507 GBV_ILN_4012 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4326 GBV_ILN_4333 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4393 GBV_ILN_4700 AR 30 2022 621-632
language	English
source	In Molecular Therapy: Nucleic Acids 30(2022), Seite 621-632 volume:30 year:2022 pages:621-632
sourceStr	In Molecular Therapy: Nucleic Acids 30(2022), Seite 621-632 volume:30 year:2022 pages:621-632
format_phy_str_mv	Article
institution	findex.gbv.de
topic_facet	MT: RNA/DNA Editing CRISPR-Cas9 primary human B cells B cell differentiation plasmablasts plasma cells Therapeutics. Pharmacology
isfreeaccess_bool	true
container_title	Molecular Therapy: Nucleic Acids
authorswithroles_txt_mv	Tuan Anh Le @@aut@@ Van Trung Chu @@aut@@ Andreia C. Lino @@aut@@ Eva Schrezenmeier @@aut@@ Christopher Kressler @@aut@@ Dania Hamo @@aut@@ Klaus Rajewsky @@aut@@ Thomas Dörner @@aut@@ Van Duc Dang @@aut@@
publishDateDaySort_date	2022-01-01T00:00:00Z
hierarchy_top_id	718632702
id	DOAJ083776613
language_de	englisch
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callnumber-first	R - Medicine
author	Tuan Anh Le
spellingShingle	Tuan Anh Le misc RM1-950 misc MT: RNA/DNA Editing misc CRISPR-Cas9 misc primary human B cells misc B cell differentiation misc plasmablasts misc plasma cells misc Therapeutics. Pharmacology Efficient CRISPR-Cas9-mediated mutagenesis in primary human B cells for identifying plasma cell regulators
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topic_title	RM1-950 Efficient CRISPR-Cas9-mediated mutagenesis in primary human B cells for identifying plasma cell regulators MT: RNA/DNA Editing CRISPR-Cas9 primary human B cells B cell differentiation plasmablasts plasma cells
topic	misc RM1-950 misc MT: RNA/DNA Editing misc CRISPR-Cas9 misc primary human B cells misc B cell differentiation misc plasmablasts misc plasma cells misc Therapeutics. Pharmacology
topic_unstemmed	misc RM1-950 misc MT: RNA/DNA Editing misc CRISPR-Cas9 misc primary human B cells misc B cell differentiation misc plasmablasts misc plasma cells misc Therapeutics. Pharmacology
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title	Efficient CRISPR-Cas9-mediated mutagenesis in primary human B cells for identifying plasma cell regulators
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title_full	Efficient CRISPR-Cas9-mediated mutagenesis in primary human B cells for identifying plasma cell regulators
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journal	Molecular Therapy: Nucleic Acids
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author_browse	Tuan Anh Le Van Trung Chu Andreia C. Lino Eva Schrezenmeier Christopher Kressler Dania Hamo Klaus Rajewsky Thomas Dörner Van Duc Dang
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title_sort	efficient crispr-cas9-mediated mutagenesis in primary human b cells for identifying plasma cell regulators
callnumber	RM1-950
title_auth	Efficient CRISPR-Cas9-mediated mutagenesis in primary human B cells for identifying plasma cell regulators
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 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. Specifically, 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.
abstractGer	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 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. Specifically, 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.
abstract_unstemmed	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 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. Specifically, 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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title_short	Efficient CRISPR-Cas9-mediated mutagenesis in primary human B cells for identifying plasma cell regulators
url	https://doi.org/10.1016/j.omtn.2022.11.016 https://doaj.org/article/174f4a9188c14eeabc505f36d39a9a14 http://www.sciencedirect.com/science/article/pii/S2162253122003080 https://doaj.org/toc/2162-2531
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Efficient CRISPR-Cas9-mediated mutagenesis in primary human B cells for identifying plasma cell regulators

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