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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Genome-wide in-locus epitope tagging of Arabidopsis proteins using prime editors

  • Cheljong Hong (Department of Chemistry, Seoul National University) ;
  • Jun Hee Han (Department of Chemistry, Hanyang University) ;
  • Gue-Ho Hwang (Research Center of Genomic Medicine Institute, Seoul National University College of Medicine) ;
  • Sangsu Bae (Research Center of Genomic Medicine Institute, Seoul National University College of Medicine) ;
  • Pil Joon Seo (Department of Chemistry, Seoul National University)
  • Received : 2023.04.11
  • Accepted : 2023.06.19
  • Published : 2024.01.31
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Abstract

Prime editors (PEs), which are CRISPR-Cas9 nickase (H840A)-reverse transcriptase fusion proteins programmed with prime editing guide RNAs (pegRNAs), can not only edit bases but also install transversions, insertions, or deletions without both donor DNA and double-strand breaks at the target DNA. As the demand for in-locus tagging is increasing, to reflect gene expression dynamics influenced by endogenous genomic contexts, we demonstrated that PEs can be used to introduce the hemagglutinin (HA) epitope tag to a target gene locus, enabling molecular and biochemical studies using in-locus tagged plants. To promote genome-wide in-locus tagging, we also implemented a publicly available database that designs pegRNAs for in-locus tagging of all the Arabidopsis genes.

Keywords

Acknowledgement

Analysis of sequencing data was mostly carried out using the computing server at the Genomic Medicine Institute Research Service Center. This research was supported by grants from the National Research Foundation of Korea (NRF) no. 2021R1AC3012908 to S.B. and no. NRF-2022R1A2B5B02001266 to P.J.S. This work was also supported by a grant from the New breeding technologies development Program (PJ01653002 to P.J.S.), Rural Development Administration.

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