Helicase-AID: A novel molecular device for base editing at random genomic loci

Jie Wang, Dongdong Zhao, Ju Li, Muzi Hu, Xiuqing Xin, Marcus A. Price, Qingyan Li, Li Liu, Siwei Li, Susan J. Rosser, Chunzhi Zhang, Changhao Bi, Xueli Zhang

Research output: Contribution to journalArticlepeer-review

Abstract / Description of output

CRISPR-enabled deaminase base editing has become a powerful tool for precisely editing nucleotides on the chromosome. In this study DNA helicases, such as Escherichia coli DnaB, were fused to activation-induced cytidine deaminase (AID) to form enzyme complexes which randomly introduces edited bases throughout the chromosome. DnaB-AID was found to increase 2.5 × 103 fold relative to the mutagenesis frequency of wildtype. 97.9% of these edits were observed on the leading strand during DNA replication suggesting deamination to be highly coordinated with DNA replication. Using DnaB-AID, a 371.4% increase in β-carotene production was obtained following four rounds of editing. In Saccharomyces cerevisiae Helicase-AID was constructed by fusing AID to one of the subunits of eukaryotic helicase Mcm2-7 complex, MCM5. Using MCM5-AID, the average editing efficiency of five strains was 2.1 ± 0.4 × 103 fold higher than the native genomic mutation rate. MCM5-AID was able to improve β-carotene production of S. cerevisiae 4742crt by 75.4% following eight rounds of editing. The S. cerevisiae MCM5-AID technique is the first biological tool for generating and accumulating single base mutations in eukaryotic chromosomes. Since the helicase complex is highly conservative in all eukaryotes, Helicase-AID could be adapted for various applications and research in all eukaryotic cells.
Original languageEnglish
Pages (from-to)396-402
Number of pages7
JournalMetabolic engineering
Early online date17 Aug 2021
Publication statusE-pub ahead of print - 17 Aug 2021

Keywords / Materials (for Non-textual outputs)

  • cytidine deaminase
  • random base editing
  • helicase


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