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

BMB Rep

Department of Chemistry, Seoul National University, Seoul 08826; Plant Genomics and Breeding Institute, Seoul National University, Seoul 08826, Korea.

Published: January 2024

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].

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10828436PMC
http://dx.doi.org/10.5483/BMBRep.2023-0055DOI Listing

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