Kai Hua1,2, Xiaoping Tao1, Liang Wei-yi1,2, Zhaoxia Zhang1,2, Runyu Gou3, Jian‐Kang Zhu4,1
1Shanghai Center for Plant Stress Biology CAS Center of Excellence in Molecular Plant Sciences Chinese Academy of Sciences Shanghai China
2University of Chinese Academy of Sciences, Beijing, China
3College of Life Sciences, Nanjing Agricultural University, Nanjing, China
4Department of Horticulture and Landscape Architecture, Purdue University, West Lafayette, IN, USA
Tóm tắt
SummaryAdenine base editors (ABEs) have been exploited to introduce targeted adenine (A) to guanine (G) base conversions in various plant genomes, including rice, wheat and Arabidopsis. However, the ABEs reported thus far are all quite inefficient at many target sites in rice, which hampers their applications in plant genome engineering and crop breeding. Here, we show that unlike in the mammalian system, a simplified base editor ABE‐P1S (Adenine Base Editor‐Plant version 1 Simplified) containing the ecTadA*7.10‐nSpCas9 (D10A) fusion has much higher editing efficiency in rice compared to the widely used ABE‐P1 consisting of the ecTadA‐ecTadA*7.10‐nSpCas9 (D10A) fusion. We found that the protein expression level of ABE‐P1S is higher than that of ABE‐P1 in rice calli and protoplasts, which may explain the higher editing efficiency of ABE‐P1S in different rice varieties. Moreover, we demonstrate that the ecTadA*7.10‐nCas9 fusion can be used to improve the editing efficiency of other ABEs containing SaCas9 or the engineered SaKKH‐Cas9 variant. These more efficient ABEs will help advance trait improvements in rice and other crops.
