title: Editing of mouse and human immunoglobulin genes by CRISPR-Cas9 system.
abstract: 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.
authors.affiliation: Department of Pathology, Children's Hospital Boston and Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Taek-Chin
authors.initials: TC
authors.lastname: Cheong
authors.affiliation: Department of Pathology, Children's Hospital Boston and Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Mara
authors.initials: M
authors.lastname: Compagno
authors.affiliation: Department of Pathology, Children's Hospital Boston and Harvard Medical School, Boston, Massachusetts 02115, USA.|Department of Molecular Biotechnology and Health Sciences, University of Torino, Torino 10126, Italy.
authors.forename: Roberto
authors.initials: R
authors.lastname: Chiarle
mesh_terms: D000818:Animals; D064113:CRISPR-Cas Systems; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D005260:Female; D006801:Humans; D017578:Immunoglobulin Class Switching; D007136:Immunoglobulins / Q000235:genetics*; D051379:Mice; D017393:RNA Editing; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000235:genetics
chemical_list: D007136:Immunoglobulins; D000094704:RNA, Guide, CRISPR-Cas Systems