TY - JOUR
T1 - Gene Editing of Primary Rhesus Macaque B Cells
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
Y1 - 2023/2/10
N2 - 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 (
AB - 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 (
UR - http://hdl.handle.net/10754/689152
UR - https://www.jove.com/t/64858/gene-editing-of-primary-rhesus-macaque-b-cells
UR - http://www.scopus.com/inward/record.url?scp=85148528489&partnerID=8YFLogxK
U2 - 10.3791/64858
DO - 10.3791/64858
M3 - Article
C2 - 36847375
SN - 1940-087X
VL - 2023
JO - Journal of Visualized Experiments
JF - Journal of Visualized Experiments
IS - 192
ER -