Cheong Taek-Chin, Compagno Mara, Chiarle Roberto
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.
Nat Commun. 2016 Mar 9;7:10934. doi: 10.1038/ncomms10934.
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.
CRISPR-Cas9系统用于编辑基因组的应用已广泛扩展，包括DNA基因敲除、缺失、染色体重排、RNA编辑和全基因组筛选。在此，我们展示了CRISPR-Cas9技术在编辑小鼠和人类免疫球蛋白（Ig）基因方面的应用。通过用逆转录病毒或慢病毒将Cas9和引导RNA（gRNA）递送至IgM(+)小鼠B细胞和杂交瘤，我们诱导IgH链向所需亚类的类别转换重组（CSR）。同样，我们在所有测试的人类B细胞系中高效诱导CSR至靶向的IgH亚类。最后，我们改造小鼠杂交瘤以分泌Fab'片段而非完整的Ig。我们的结果表明，小鼠和人类细胞中的Ig基因可以被编辑，以获得任何有助于研究正常和淋巴瘤B细胞生物学的所需IgH转换。我们还提出了可能改变抗体生产技术的应用。
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