Engineered Un1Cas12f1 for multiplex genome editing with enhanced activity and targeting scope

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Author notes

1. These authors contributed equally: Yanan Huo, Jiale Mei, Dan Zhang.

Authors and Affiliations

1. Shanghai Frontiers Science Center of Genome Editing and Cell Therapy, Shanghai Key Laboratory of Regulatory Biology, School of Life Sciences, East China Normal University, Shanghai, China

   Yanan Huo, Jiale Mei, Dan Zhang, Bing Yan, Chao Dong, Shuming Yin, Meizhen Liu, Xinyan Wang, Dan Chen, Yuting Guan, Gaojie Song, Bing Du, Dali Li & Liren Wang

2. Department of Urology, Children’s Hospital of Fudan University, Shanghai, China

   Dexin Zhang

3. Shanghai Engineering Research Center of Industrial Microorganisms, Shanghai, China

   Yongming Wang

4. Department of Biomedical Sciences and Tung Biomedical Sciences Centre, City University of Hong Kong, Kowloon, Hong Kong, SAR, China

   Zongli Zheng

5. Department of Dermatology, Xiangya Hospital & School of Life Sciences & Furong Laboratory, Central South University, Changsha, China

   Hong Liu

6. Shanghai Academy of Natural Sciences (SANS), Shanghai, China

   Dali Li

7. Hangzhou Institute of Medicine (HIM), Chinese Academy of Sciences, Hangzhou, 310018, China

   Lei Yang

Contributions

Y.H., L.W., L.Y., and D.L. conceived the rational engineering of Un1Cas12f1. Y.H., L.W., and D.C. performed the experiments of Un1Cas12f1 mutation library construction. Y.H., L.W., and J.M. performed the experiments of bacterial positive selection assay. Y.H., J.M., B.Y. performed the experiments of plasmid construction, cell culture, cell transfection, cell sorting, and genomic DNA preparation. Y. H. and J. M. performed the experiments of PCR, reverse transcription, qPCR and prepared the HTS libraries. L.W., X.W., and Dan Zhang performed computational analysis. Y.H., C.D., and S.Y. performed the experiments of lentiviral vector production and the creation of stable cell lines. Y.H., J.M., Dexin Zhang and M.L. performed the animal experiments. G.S. performed the structural analysis. L.W., Y.H., L.Y., D.L., Y.G., B.D., Y.W., H.L., and Z.Z. designed the experiments and wrote the manuscript with the input from all the authors. D.L., L.Y., and L.W. supervised the study.

Corresponding authors

Correspondence to Dali Li, Lei Yang or Liren Wang.

Competing interests

D.L., L.W, Y.H., J.M, L.Y, and Dan Zhang have submitted patent applications (application number CN 202510015341.4, under review) based on the results reported in this study. This patent mainly relates to evoCas12f in this paper. The remaining authors declare no competing interests.

Peer review information

Nature Communications thanks the anonymous reviewers for their contribution to the peer review of this work. A peer review file is available.

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