Genome-wide in-locus epitope tagging of Arabidopsis proteins using prime editors

Cheljong Hong; Jun Hee Han; Gue-Ho Hwang; Sangsu Bae; Pil Joon Seo

doi:10.5483/BMBRep.2023-0055

Genome-wide in-locus epitope tagging of Arabidopsis proteins using prime editors

BMB Rep. 2024 Jan;57(1):66-70. doi: 10.5483/BMBRep.2023-0055.

Authors

Cheljong Hong¹, Jun Hee Han², Gue-Ho Hwang³, Sangsu Bae⁴, Pil Joon Seo⁵

Affiliations

¹ Department of Chemistry, Seoul National University, Seoul 08826, Korea.
² Department of Chemistry, Hanyang University, Seoul 04673, Korea.
³ Medical Research Center of Genomic Medicine Institute, Seoul National University College of Medicine, Seoul 03080, Korea.
⁴ Medical Research Center of Genomic Medicine Institute, Seoul National University College of Medicine, Seoul 03080; Cancer Research Institute, Seoul National University College of Medicine, Seoul 03080; Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul 03080, Korea.
⁵ Department of Chemistry, Seoul National University, Seoul 08826; Plant Genomics and Breeding Institute, Seoul National University, Seoul 08826, Korea.

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. [BMB Reports 2024; 57(1): 66-70].

Publication types

News

MeSH terms

Arabidopsis Proteins* / genetics
CRISPR-Cas Systems* / genetics
DNA / genetics
Gene Editing
RNA, Guide, CRISPR-Cas Systems

Substances

RNA, Guide, CRISPR-Cas Systems
Arabidopsis Proteins
DNA

Grants and funding

ACKNOWLEDGEMENTS 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.