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Editing of mouse and human immunoglobulin genes by CRISPR-Cas9 system

Applications of the CRISPR-Cas9 system to edit the genome have widely expanded to include DNA gene knock-out, deletions, chromosomal rearrangements, RNA editing and genome-wide screenings. Here we show the application of CRISPR-Cas9 technology to edit the mouse and human immunoglobulin (Ig) genes. B...

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Autores principales: Cheong, Taek-Chin, Compagno, Mara, Chiarle, Roberto
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4786874/
https://www.ncbi.nlm.nih.gov/pubmed/26956543
http://dx.doi.org/10.1038/ncomms10934
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author Cheong, Taek-Chin
Compagno, Mara
Chiarle, Roberto
author_facet Cheong, Taek-Chin
Compagno, Mara
Chiarle, Roberto
author_sort Cheong, Taek-Chin
collection PubMed
description Applications of the CRISPR-Cas9 system to edit the genome have widely expanded to include DNA gene knock-out, deletions, chromosomal rearrangements, RNA editing and genome-wide screenings. Here we show the application of CRISPR-Cas9 technology to edit the mouse and human immunoglobulin (Ig) genes. By delivering Cas9 and guide-RNA (gRNA) with retro- or lenti-virus to IgM(+) mouse B cells and hybridomas, we induce class-switch recombination (CSR) of the IgH chain to the desired subclass. Similarly, we induce CSR in all human B cell lines tested with high efficiency to targeted IgH subclass. Finally, we engineer mouse hybridomas to secrete Fab′ fragments instead of the whole Ig. Our results indicate that Ig genes in mouse and human cells can be edited to obtain any desired IgH switching helpful to study the biology of normal and lymphoma B cells. We also propose applications that could transform the technology of antibody production.
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spelling pubmed-47868742016-03-16 Editing of mouse and human immunoglobulin genes by CRISPR-Cas9 system Cheong, Taek-Chin Compagno, Mara Chiarle, Roberto Nat Commun Article Applications of the CRISPR-Cas9 system to edit the genome have widely expanded to include DNA gene knock-out, deletions, chromosomal rearrangements, RNA editing and genome-wide screenings. Here we show the application of CRISPR-Cas9 technology to edit the mouse and human immunoglobulin (Ig) genes. By delivering Cas9 and guide-RNA (gRNA) with retro- or lenti-virus to IgM(+) mouse B cells and hybridomas, we induce class-switch recombination (CSR) of the IgH chain to the desired subclass. Similarly, we induce CSR in all human B cell lines tested with high efficiency to targeted IgH subclass. Finally, we engineer mouse hybridomas to secrete Fab′ fragments instead of the whole Ig. Our results indicate that Ig genes in mouse and human cells can be edited to obtain any desired IgH switching helpful to study the biology of normal and lymphoma B cells. We also propose applications that could transform the technology of antibody production. Nature Publishing Group 2016-03-09 /pmc/articles/PMC4786874/ /pubmed/26956543 http://dx.doi.org/10.1038/ncomms10934 Text en Copyright © 2016, Nature Publishing Group, a division of Macmillan Publishers Limited. All Rights Reserved. http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Cheong, Taek-Chin
Compagno, Mara
Chiarle, Roberto
Editing of mouse and human immunoglobulin genes by CRISPR-Cas9 system
title Editing of mouse and human immunoglobulin genes by CRISPR-Cas9 system
title_full Editing of mouse and human immunoglobulin genes by CRISPR-Cas9 system
title_fullStr Editing of mouse and human immunoglobulin genes by CRISPR-Cas9 system
title_full_unstemmed Editing of mouse and human immunoglobulin genes by CRISPR-Cas9 system
title_short Editing of mouse and human immunoglobulin genes by CRISPR-Cas9 system
title_sort editing of mouse and human immunoglobulin genes by crispr-cas9 system
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4786874/
https://www.ncbi.nlm.nih.gov/pubmed/26956543
http://dx.doi.org/10.1038/ncomms10934
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