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Improving the Efficiency of Prime Editing with Epegrnas and High-Temperature Treatment in Rice
Jinpeng Zou, Xiangbing Meng, Qing Liu, Meiqi Shang, Kejian Wang, Jiayang Li, Hong Yu, Chun Wang
SCIENCE CHINA Life Sciences
Abstract
Prime editing (PE) systems are important genome-editing tools developed for mediating arbitrary small DNA insertions, deletions and all 12 base-to-base conversions using a CRISPR-nCas9 fusion of an engineered M-MLV and PE guide RNAs (pegRNAs) (Anzalone et al., 2019). At present, PE systems are applied in many plants; however, compared with the PE efficiency of 20%–70% in mammalian cells, the PE efficiency in plant cells is low, which limits the application in breeding. Several studies have reported that PE efficiencies in plants could be improved by optimising the length of the primer binding site (PBS), length of the RT template and position of the nicking single guide RNA (sgRNA). Other options include the use of different promoters and performing N-terminal M-MLV-nCas9 fusion. However, the current PE efficiencies are less than satisfactory in plants (Jiang et al., 2020; Li et al., 2020; Lin et al., 2020; Lu et al., 2021; Tang et al., 2020; Xu et al., 2020a; Xu et al., 2022). Recently, a study on mammalian cells showed that incorporating the structured RNA motifs into the 3′ terminus of pegRNAs could reduce the degradation of pegRNAs and significantly improve the PE efficiencies by 3–4 fold (Nelson et al., 2022). This study prompted us to consider whether the engineered pegRNAs (epegRNAs) could enhance the PE efficiencies in rice.
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论文编号: |
DOI:10.1007/s11427-022-2147-2 |
论文题目: |
Improving the Efficiency of Prime Editing with Epegrnas and High-Temperature Treatment in Rice |
英文论文题目: |
Improving the Efficiency of Prime Editing with Epegrnas and High-Temperature Treatment in Rice |
第一作者: |
Jinpeng Zou, Xiangbing Meng, Qing Liu, Meiqi Shang, Kejian Wang, Jiayang Li, Hong Yu, Chun Wang |
英文第一作者: |
Jinpeng Zou, Xiangbing Meng, Qing Liu, Meiqi Shang, Kejian Wang, Jiayang Li, Hong Yu, Chun Wang |
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2022-06-30 |
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摘要: |
Prime editing (PE) systems are important genome-editing tools developed for mediating arbitrary small DNA insertions, deletions and all 12 base-to-base conversions using a CRISPR-nCas9 fusion of an engineered M-MLV and PE guide RNAs (pegRNAs) (Anzalone et al., 2019). At present, PE systems are applied in many plants; however, compared with the PE efficiency of 20%–70% in mammalian cells, the PE efficiency in plant cells is low, which limits the application in breeding. Several studies have reported that PE efficiencies in plants could be improved by optimising the length of the primer binding site (PBS), length of the RT template and position of the nicking single guide RNA (sgRNA). Other options include the use of different promoters and performing N-terminal M-MLV-nCas9 fusion. However, the current PE efficiencies are less than satisfactory in plants (Jiang et al., 2020; Li et al., 2020; Lin et al., 2020; Lu et al., 2021; Tang et al., 2020; Xu et al., 2020a; Xu et al., 2022). Recently, a study on mammalian cells showed that incorporating the structured RNA motifs into the 3′ terminus of pegRNAs could reduce the degradation of pegRNAs and significantly improve the PE efficiencies by 3–4 fold (Nelson et al., 2022). This study prompted us to consider whether the engineered pegRNAs (epegRNAs) could enhance the PE efficiencies in rice. |
英文摘要: |
Prime editing (PE) systems are important genome-editing tools developed for mediating arbitrary small DNA insertions, deletions and all 12 base-to-base conversions using a CRISPR-nCas9 fusion of an engineered M-MLV and PE guide RNAs (pegRNAs) (Anzalone et al., 2019). At present, PE systems are applied in many plants; however, compared with the PE efficiency of 20%–70% in mammalian cells, the PE efficiency in plant cells is low, which limits the application in breeding. Several studies have reported that PE efficiencies in plants could be improved by optimising the length of the primer binding site (PBS), length of the RT template and position of the nicking single guide RNA (sgRNA). Other options include the use of different promoters and performing N-terminal M-MLV-nCas9 fusion. However, the current PE efficiencies are less than satisfactory in plants (Jiang et al., 2020; Li et al., 2020; Lin et al., 2020; Lu et al., 2021; Tang et al., 2020; Xu et al., 2020a; Xu et al., 2022). Recently, a study on mammalian cells showed that incorporating the structured RNA motifs into the 3′ terminus of pegRNAs could reduce the degradation of pegRNAs and significantly improve the PE efficiencies by 3–4 fold (Nelson et al., 2022). This study prompted us to consider whether the engineered pegRNAs (epegRNAs) could enhance the PE efficiencies in rice. |
刊物名称: |
SCIENCE CHINA Life Sciences |
英文刊物名称: |
SCIENCE CHINA Life Sciences |
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其它备注: |
Jinpeng Zou, Xiangbing Meng, Qing Liu, Meiqi Shang, Kejian Wang, Jiayang Li, Hong Yu, Chun Wang. Improving the Efficiency of Prime Editing with Epegrnas and High-Temperature Treatment in Rice. SCIENCE CHINA Life Sciences. DOI:10.1007/s11427-022-2147-2 |
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