Protocol for genetic dissection of class switch recombination using genome-editing tools

Protocol for genetic dissection of class switch recombination using genome-editing tools

Summary: Antibody class switch recombination is achieved through programmed DNA damage, and the processing of programmed DNA lesions requires the coordinated action of many DNA metabolic factors. Here, we present a protocol for inducing class switch recombination using base editors or CRISPR-Cas9. W...

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Bibliographic Details
Main Authors: Pengfei Dai, Tingting Liu, Dingpeng Yang, Yifeng Luo, Liu Daisy Liu, Xia Xie, Yafang Shang, Xiaojing Liu, Fei-Long Meng
Format: Article
Language:English
Published: Elsevier 2025-06-01
Series:STAR Protocols
Subjects:
cell-based assays
immunology
CRISPR
Online Access:http://www.sciencedirect.com/science/article/pii/S2666166725002886
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Summary:Summary: Antibody class switch recombination is achieved through programmed DNA damage, and the processing of programmed DNA lesions requires the coordinated action of many DNA metabolic factors. Here, we present a protocol for inducing class switch recombination using base editors or CRISPR-Cas9. We provide optimized guide RNA (gRNA) sequences and describe steps for cytokine activation, electroporation, surface immunoglobulin detection, and data analysis. This method allows researchers to investigate the involvement of specific factors in antibody diversification and elucidate their functional roles.For complete details on the use and execution of this protocol, please refer to Dai et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
ISSN:2666-1667

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