Efficient generation of homozygous substitutions in rice in one generation utilizing an rABE8e base editor

• Published in: Journal of integrative plant biology Vol. 63; no. 9; pp. 1595 - 1599
• Main Authors: Wei, Chuang, Wang, Chong, Jia, Meng, Guo, Hong‐Xuan, Luo, Peng‐Yu, Wang, Mu‐Gui, Zhu, Jian‐Kang, Zhang, Hui
• Format: Journal Article
• Language: English
• Published: China (Republic : 1949- ) Wiley Subscription Services, Inc 01.09.2021; Shanghai Key Laboratory of Plant Molecular Sciences,College of Life Sciences,Shanghai Normal University,Shanghai 200234,China%Shanghai Center for Plant Stress Biology and Center for Excellence in Molecular Plant Sciences,The Chinese Academy of Sciences,Shanghai 200032,China; Development Center of Plant Germplasm Resources,College of Life Sciences,Shanghai Normal University,Shanghai 200234,China
• Subjects: Adenine; Aminohydrolases - metabolism; base editing; CRISPR; CRISPR-Associated Protein 9; Deamination; Deoxyribonucleic acid; DNA; Editing; Gene Editing - methods; genes; homozygosity; Optimization; Oryza - genetics; plant biology; rABE8e; rice; CRISPR; rABE8e; rice; base editing
• ISSN: 1672-9072; 1744-7909; 1744-7909
• DOI: 10.1111/jipb.13089

Summary A new deaminase, TadA8e, was recently evolved in the laboratory. TadA8e catalyzes DNA deamination over 1,000 times faster than ABE7.10. We developed a high‐efficiency adenine base editor, rABE8e (rice ABE8e), combining monomeric TadA8e, bis‐bpNLS and codon optimization. rABE8e had substantially increased editing efficiencies at NG‐protospacer adjacent motif (PAM) and NGG‐PAM target sequences compared with ABEmax. For most targets, rABE8e exhibited nearly 100% editing efficiency and high homozygous substitution rates in the specific editing window, especially at Positions A5 and A6. The ability to rapidly generate plant materials with homozygous base substitutions will benefit gene function research and precision molecular breeding. The high‐efficiency adenine base editor rABE8e combines monomeric TadA8e, bis‐bpNLS and codon optimization. rABE8e exhibited close to 100% editing efficiency and high homozygous substitution rates. With the advent of rABE8e, it is now easy to obtain homozygous substitution rice materials in the T0 generation.