554 / 2019-03-15 21:22:54

Precision Genome Engineering through Adenine Base Editing in Plants

CRISPR,Adenine base editing,ABE,FT,PDS3,neofunctionalization,mis-splicing

全体主题 > Omics& New Technology

CRISPR genome editing holds promise as a revolutionary tool for basic research and biotechnology. Canonical CRISPR nucleases cleave DNA in a targeted manner resulting in small indels at target sites. However, inducing point mutations rather than indels remains a challenge, although point mutagenesis is one of the key strategies to decode or create genomic variations and furthermore to achieve crop improvement. The recent development of adenine base editors (ABEs), composed of the Cas9 nickase and engineered tRNA adenosine deaminases, has enabled efficient programmable A/T-to-G/C base conversions in eukaryotic cells. For in planta ABE applications, we developed a plant-compatible ABE system that can be successfully applied to Arabidopsis plants to obtain organisms with desired phenotypes with germline transmission. For example, targeted precise A-to-G substitutions generated a single amino-acid change in the FT protein or mis-splicing of the PDS3 RNA transcript with germline transmission of such edited alleles, and we could thereby obtain transgenic plants with late flowering and albino phenotypes, respectively. We demonstrate "proof-of-concept" in planta ABE applications that can lead to induced protein neo-functionalization or mRNA mis-splicing, opening up new avenues for plant genome engineering and biotechnology.

- Published at Nature Plants 2018 (also featured in News & Views in the issue, and the figure used as hero image in the Nature Plants website)-