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.

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