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Mutagenesis system by fusing cytidine deaminase with T7 promoter in yeast
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  • Zi-rui Huang,
  • Bing-Zhi Li,
  • Xiang-rong Chen,
  • Dan-feng Liu,
  • You-zhi Cui,
  • Yingjin Yuan
Zi-rui Huang
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Bing-Zhi Li
Tianjin University

Corresponding Author:[email protected]

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Xiang-rong Chen
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Dan-feng Liu
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You-zhi Cui
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Yingjin Yuan
School of Chemical Engineering and Technology
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The occurrence of random mutations can increase the diversity of the genome and promote the evolutionary process of organisms. High efficiency mutagenesis techniques significantly accelerate the evolutionary process. In this work, we describe a targeted in vivo mutagenesis system to significantly increase mutation frequency and generate mutations across all four nucleotides. We constructed different DNA-modifying enzyme-PmCDA1-T7 RNA polymerase fusion proteins, achieved targeted mutagenesis by flanking the target gene with T7 promoters, and tuned the mutation spectra by introducing different DNA-modifying enzymes. With the mutagenesis fusion proteins, the mutation frequency of the target gene could reach 5.13x10-3, and the proportion of non-C→T mutations is 10~11-fold higher than the cytidine-based evolutionary tools. We also demonstrated that our mutagenesis tools could be used to evolve the essential enzyme in the β-carotene biosynthesis process and generate mutations with different types.
26 Mar 2023Submitted to Biotechnology Journal
27 Mar 2023Submission Checks Completed
27 Mar 2023Assigned to Editor
28 Mar 2023Reviewer(s) Assigned
18 May 2023Review(s) Completed, Editorial Evaluation Pending
19 May 2023Editorial Decision: Revise Major
26 Jun 20231st Revision Received
26 Jun 2023Submission Checks Completed
26 Jun 2023Assigned to Editor
26 Jun 2023Reviewer(s) Assigned
12 Jul 2023Review(s) Completed, Editorial Evaluation Pending
12 Jul 2023Editorial Decision: Revise Major
20 Jul 20232nd Revision Received
20 Jul 2023Submission Checks Completed
20 Jul 2023Assigned to Editor
20 Jul 2023Reviewer(s) Assigned
25 Jul 2023Review(s) Completed, Editorial Evaluation Pending
28 Jul 2023Editorial Decision: Accept