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Genome-wide In-Locus Epitope Tagging of Arabidopsis Proteins using Prime Editors
Pil Joon Seo 1,6,* (Associate Professor), Sangsu Bae 3,4,5 (Associate Professor), Cheljong Hong 1,# (Graduate student), Jun Hee Han 2,# (Graduate student), Gue-Ho Hwang 3 (Postdoc researcher)
1Department of Chemistry, Seoul National University, Seoul 08826, Republic of Korea,
2Department of Chemistry, Hanyang University, Seoul 04673, Republic of Korea,
3Medical Research Center of Genomic Medicine Institute and 4Cancer Research Institute and 5Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul 03080, Republic of Korea,
6Plant Genomics and Breeding Institute, Seoul National University, Seoul 08826, Republic of 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 target DNA. As the demands for in-locus tagging are 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 Arabidopsis genes.
Abstract, Accepted Manuscript(in press) [Submitted on April 14, 2023, Accepted on June 19, 2023]
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