This study is led by Professor Wen Wang (School of Ecology and Environment, Northwestern Polytechnical University, Xi'an, China). The authors tested three newly identified TnpB proteins: ISAam1 (369 aa), ISYmu1 (382 aa), and ISDra2 (408 aa) in rice to investigate the ability of TnpB transposon endonucleases for editing target genes in plants for the first time. The authors first constructing TnpB plant genome editing vectors that targeting OsPDS to evaluate the genome editing efficiency of the three TnpB sytems in stable transgenic rice. Subsequently, the authors targeting OsYSA and OsCERK1 to further evaluate the genome editing efficiency of the three TnpB sytems in rice callus.
Plant genome editing vectors were constructed to target OsPDS first. Plants phenotypic analysis and sequencing results showed that ISDra2 and ISYmu1 TnpB systems could edit plant genome, while no editing was found in ISAam1 system. Meanwhile, whole genome sequencing analysis of albino seedlings targeting OsPDS did not detect any off-target mutation. The other two target genes, OsYSA and OsCERK1, were then tested in rice callus. NGS analysis results showed that both target genes were edited at the target site in ISDra2 and ISYmu1 TnpB systems, but no mutation were fund in ISAam1 system, which was consistent with the results in plants.
By testing three TnpB systems editing targeting three endogenous genes in rice, the authors evaluate the gene editing efficiency and accuracy of the three TnpB systems in rice. In summary, this work demonstrate for the first time the successful TnpB-mediated gene editing in plants and show that the ISDra2 and ISYmu1 nucleases possess considerable gene editing efficiency, which laid the groundwork for the development of additional hypercompact plant genome editing tools.
See the article:
Genome editing in plants using the TnpB transposase system
https://link.springer.com/article/10.1007/s42994-024-00172-6
