Open AccessCRISPR RNA ‐guided integrase enables high‐efficiency targeted genome engineering in Agrobacterium tumefaciens
Author(s) -
Aliu Ephraim,
Lee Keunsub,
Wang Kan
Publication year - 2022
Publication title -
plant biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.525
H-Index - 115
eISSN - 1467-7652
pISSN - 1467-7644
DOI - 10.1111/pbi.13872
Subject(s) - agrobacterium tumefaciens , biology , integrase , agrobacterium , crispr , genome engineering , genome , gene , transfer dna , genetics , computational biology , dna , genome editing , transformation (genetics)
Summary Agrobacterium tumefaciens , the causal agent of plant crown gall disease, has been widely used to genetically transform many plant species. The inter‐kingdom gene transfer capability made Agrobacterium an essential tool and model system to study the mechanism of exporting and integrating a segment of bacterial DNA into the plant genome. However, many biological processes such as Agrobacterium ‐host recognition and interaction are still elusive. To accelerate the understanding of this important plant pathogen and further improve its capacity in plant genetic engineering, we adopted a CRISPR RNA‐guided integrase system for Agrobacterium genome engineering. In this work, we demonstrate that IN sertion of T ransposable E lements by G uide R NA– A ssisted T arg E ting (INTEGRATE) can efficiently generate DNA insertions to enable targeted gene knockouts. In addition, in conjunction with Cre‐ loxP recombination system, we achieved precise deletions of large DNA fragments. This work provides new genetic engineering strategies for Agrobacterium species and their gene functional analyses.