The robust, high-throughput, and temporally regulated roxCre and loxCre reporting systems for genetic modifications in vivo

Author:

Shi Mengyang1,Li Jie1,Liu Xiuxiu1,Liu Kuo2,Pu Wenjuan1,Weng Wendong1,Zhang Shaohua1,Zhao Huan1,Lui Kathy O.3ORCID,Zhou Bin124ORCID

Affiliation:

1. New Cornerstone Science Laboratory, State Key Laboratory of Cell Biology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, University of Chinese Academy of Sciences

2. Key Laboratory of Systems Health Science of Zhejiang Province, School of Life Science, Hangzhou Institute for Advanced Study, University of Chinese Academy of Sciences

3. Department of Chemical Pathology; and Li Ka Shing Institute of Health Sciences, The Chinese University of Hong Kong, Prince of Wales Hospital

4. School of Life Science and Technology, ShanghaiTech University

Abstract

Cre-loxP technology, a cornerstone in fate mapping and in vivo gene function studies, faces challenges in achieving precise and efficient conditional mutagenesis through inducible systems. In this study, we introduce two innovative genetic tools designed to overcome these limitations. The first, loxCre, facilitates conditional gene targeting by allowing any CreER line to induce Cre expression with significantly enhanced efficiency. The second, roxCre, enables DreER-mediated Cre release, paving the way for intersectional genetic manipulation that permits both increased precision and efficiency. Both tools incorporate a fluorescent reporter for genetic lineage tracing, revealing efficient gene knockout in cells marked by the reporter simultaneously. These strategies hold great potential for precise and efficient exploration of lineage-specific gene functions, marking a significant advancement in genetic research methodologies.

Publisher

eLife Sciences Publications, Ltd

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