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Transient expression of a TaGRF4-TaGIF1 complex stimulates wheat regeneration and improves genome editing 被引量:7
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作者 fengti qiu Sinian Xing +4 位作者 Chenxiao Xue Jinxing Liu Kunling Chen Tuanyao Chai Caixia Gao 《Science China(Life Sciences)》 SCIE CAS CSCD 2022年第4期731-738,共8页
Genome editing is an unprecedented technological breakthrough but low plant regeneration frequencies and genotype dependence hinder its implementation for crop improvement. Here, we found that transient expression of ... Genome editing is an unprecedented technological breakthrough but low plant regeneration frequencies and genotype dependence hinder its implementation for crop improvement. Here, we found that transient expression of a complex of the growth regulators TaGRF4 and TaGIF1(TaGRF4-TaGIF1) increased regeneration and genome editing frequency in wheat. When we introduced synonymous mutation in the miR396 target site of TaGRF4, the resulting complex(mTaGRF4-TaGIF1) performed better than original TaGRF4-TaGIF1. Use of m TaGRF4-TaGIF1 together with a cytosine base editor targeting TaALS resulted in 2-9-fold increases in regeneration and transgene-free genome editing in 11 elite common wheat cultivars. Therefore, m TaGRF4-TaGIF1 will undoubtedly be of great value in crop improvement and especially in commercial applications, since it greatly increased the range of cultivars available for transformation. 展开更多
关键词 mTaGRF4-TaGIF1 WHEAT REGENERATION genome editing transient expression
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Shortening the sgRNA-DNA interface enables SpCas9 and eSpCas9(1.1)to nick the target DNA strand 被引量:4
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作者 Rong Fan Zhuangzhuang Chai +7 位作者 Sinian Xing Kunling Chen fengti qiu Tuanyao Chai Jin-Long qiu Zhengbin Zhang Huawei Zhang Caixia Gao 《Science China(Life Sciences)》 SCIE CAS CSCD 2020年第11期1619-1630,共12页
The length of the sgRNA-DNA complementary sequence is a key factor influencing the cleavage activity of Streptococcus pyogenes Cas9(SpCas9)and its variants.The detailed mechanism remains unknown.Here,based on in vitro... The length of the sgRNA-DNA complementary sequence is a key factor influencing the cleavage activity of Streptococcus pyogenes Cas9(SpCas9)and its variants.The detailed mechanism remains unknown.Here,based on in vitro cleavage assays and base editing analysis,we demonstrate that reducing the length of this complementary region can confer nickase activity on SpCas9 and eSpCas9(1.1).We also show that these nicks are made on the target DNA strand.These properties encouraged us to develop a dual-functional system that simultaneously carries out double-strand DNA cleavage and C-to-T base conversions at separate targets.This system provides a novel tool for achieving trait stacking in plants. 展开更多
关键词 SpCas9 eSpCas9(1.1) truncated spacer DSB nickase co-editing
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