Gene Editing of Primary Rhesus Macaque B Cells

Harald Hartweger; Rajeev Gautam; Yoshiaki Nishimura; Fabian Schmidt; Kai-Hui Yao; Amelia Escolano; Mila Jankovic; Malcolm A. Martin; Michel C. Nussenzweig

doi:10.3791/64858

Gene Editing of Primary Rhesus Macaque B Cells

Harald Hartweger, Rajeev Gautam, Yoshiaki Nishimura, Fabian Schmidt, Kai-Hui Yao, Amelia Escolano, Mila Jankovic, Malcolm A. Martin, Michel C. Nussenzweig

Research output: Contribution to journal › Article › peer-review

3 Scopus citations

Abstract

B cells and their progeny are the sources of highly expressed antibodies. Their high protein expression capabilities together with their abundance, easy accessibility via peripheral blood, and amenability to simple adoptive transfers have made them an attractive target for gene editing approaches to express recombinant antibodies or other therapeutic proteins. The gene editing of mouse and human primary B cells is efficient, and mouse models for in vivo studies have shown promise, but feasibility and scalability for larger animal models have so far not been demonstrated. We, therefore, developed a protocol to edit rhesus macaque primary B cells in vitro to enable such studies. We report conditions for in vitro culture and gene-editing of primary rhesus macaque B cells from peripheral blood mononuclear cells or splenocytes using CRISPR/Cas9. To achieve the targeted integration of large (

Original language	English (US)
Journal	Journal of Visualized Experiments
Volume	2023
Issue number	192
DOIs	https://doi.org/10.3791/64858
State	Published - Feb 10 2023

Bibliographical note

KAUST Repository Item: Exported on 2023-03-01
Acknowledgements: We would like to thank Harry B. Gristick and Pamela Bjorkman for providing the RC1 antigen and the entire Nussenzweig and Martin laboratories for critical discussion. This work was supported by The Bill and Melinda Gates Foundation grant INV-002777 (to M.C.N.) and the Intramural Research Program of the National Institute of Allergy and Infectious Diseases, National Institutes of Health. (R.G. and M.A.M). M.C.N. is an HHMI Investigator.

ASJC Scopus subject areas

General Biochemistry, Genetics and Molecular Biology
General Medicine
General Immunology and Microbiology
General Chemical Engineering
General Neuroscience

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10.3791/64858

https://repository.kaust.edu.sa/bitstream/10754/689152/1/29e30c31-bbf4-4974-a810-ee8c5d4402f3.pdf

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title = "Gene Editing of Primary Rhesus Macaque B Cells",

abstract = "B cells and their progeny are the sources of highly expressed antibodies. Their high protein expression capabilities together with their abundance, easy accessibility via peripheral blood, and amenability to simple adoptive transfers have made them an attractive target for gene editing approaches to express recombinant antibodies or other therapeutic proteins. The gene editing of mouse and human primary B cells is efficient, and mouse models for in vivo studies have shown promise, but feasibility and scalability for larger animal models have so far not been demonstrated. We, therefore, developed a protocol to edit rhesus macaque primary B cells in vitro to enable such studies. We report conditions for in vitro culture and gene-editing of primary rhesus macaque B cells from peripheral blood mononuclear cells or splenocytes using CRISPR/Cas9. To achieve the targeted integration of large (",

author = "Harald Hartweger and Rajeev Gautam and Yoshiaki Nishimura and Fabian Schmidt and Kai-Hui Yao and Amelia Escolano and Mila Jankovic and Martin, {Malcolm A.} and Nussenzweig, {Michel C.}",

note = "KAUST Repository Item: Exported on 2023-03-01 Acknowledgements: We would like to thank Harry B. Gristick and Pamela Bjorkman for providing the RC1 antigen and the entire Nussenzweig and Martin laboratories for critical discussion. This work was supported by The Bill and Melinda Gates Foundation grant INV-002777 (to M.C.N.) and the Intramural Research Program of the National Institute of Allergy and Infectious Diseases, National Institutes of Health. (R.G. and M.A.M). M.C.N. is an HHMI Investigator.",

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AU - Hartweger, Harald

AU - Gautam, Rajeev

AU - Nishimura, Yoshiaki

AU - Schmidt, Fabian

AU - Yao, Kai-Hui

AU - Escolano, Amelia

AU - Jankovic, Mila

AU - Martin, Malcolm A.

AU - Nussenzweig, Michel C.

N1 - KAUST Repository Item: Exported on 2023-03-01 Acknowledgements: We would like to thank Harry B. Gristick and Pamela Bjorkman for providing the RC1 antigen and the entire Nussenzweig and Martin laboratories for critical discussion. This work was supported by The Bill and Melinda Gates Foundation grant INV-002777 (to M.C.N.) and the Intramural Research Program of the National Institute of Allergy and Infectious Diseases, National Institutes of Health. (R.G. and M.A.M). M.C.N. is an HHMI Investigator.

PY - 2023/2/10

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Gene Editing of Primary Rhesus Macaque B Cells

Abstract

Bibliographical note

ASJC Scopus subject areas

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