Multiplexed CRISPR/CAS9‐mediated engineering of pre‐clinical mouse models bearing native human B cell receptors
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44318_2020_Article_BFEMBJ2020105926.pdf
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Author(s)
Wang, Xuesong
Ray, Rashmi
Kratochvil, Sven
Melzi, Eleonora
Lin, Ying‐Cing
Giguere, Sophie
Xu, Liling
Warner, John
Cheon, Diane
Liguori, Alessia
Date Issued
December 1, 2020
Journal
The Embo Journal
Publisher
Nature Publishing Group UK
Citation
The EMBO Journal. 2020 Dec 01;40(2):EMBJ2020105926
Version
Final published version
Abstract
B‐cell receptor (BCR) knock‐in (KI) mouse models play an important role in vaccine development and fundamental immunological studies. However, the time required to generate them poses a bottleneck. Here we report a one‐step CRISPR/Cas9 KI methodology to combine the insertion of human germline immunoglobulin heavy and light chains at their endogenous loci in mice. We validate this technology with the rapid generation of three BCR KI lines expressing native human precursors, instead of computationally inferred germline sequences, to HIV broadly neutralizing antibodies. We demonstrate that B cells from these mice are fully functional: upon transfer to congenic, wild type mice at controlled frequencies, such B cells can be primed by eOD‐GT8 60mer, a germline‐targeting immunogen currently in clinical trials, recruited to germinal centers, secrete class‐switched antibodies, undergo somatic hypermutation, and differentiate into memory B cells. KI mice expressing functional human BCRs promise to accelerate the development of vaccines for HIV and other infectious diseases.
MIT Department
Ragon Institute of MGH, MIT and Harvard
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Creative Commons Attribution
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DOI of Published Version
https://doi.org/10.15252/embj.2020105926
