Nature Biotechnology
Abstract
Although prime editors (PEs) have the potential to facilitate precise genome editing in therapeutic, agricultural and research applications, their specificity has not been comprehensively evaluated. To provide a systematic assessment in plants, we first examined the mismatch tolerance of PEs in plant cells and found that the editing frequency was influenced by the number and location of mismatches in the primer binding site and spacer of the prime editing guide RNA (pegRNA). Assessing the activity of 12 pegRNAs at 179 predicted off-target sites, we detected only low frequencies of off-target edits (0.00~0.23%). Whole-genome sequencing of 29 PE-treated rice plants confirmed that PEs do not induce genome-wide pegRNA-independent off-target single-nucleotide variants or small insertions/deletions. We also show that ectopic expression of the Moloney murine leukemia virus reverse transcriptase as part of the PE does not change retrotransposon copy number or telomere structure or cause insertion of pegRNA or messenger RNA sequences into the genome.
| 论文编号: | DOI:10.1038/s41587-021-00891-x |
| 论文题目: | Genome-Wide Specificity of Prime Editors in Plants |
| 英文论文题目: | Genome-Wide Specificity of Prime Editors in Plants |
| 第一作者: | Shuai Jin, Qiupeng Lin, Yingfeng Luo, Zixu Zhu, Guanwen Liu, Yunjia Li, Kunling Chen, Jin-Long Qiu and Caixia Gao |
| 英文第一作者: | Shuai Jin, Qiupeng Lin, Yingfeng Luo, Zixu Zhu, Guanwen Liu, Yunjia Li, Kunling Chen, Jin-Long Qiu and Caixia Gao |
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| 发表年度: | 2021-04-16 |
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| 摘要: | Although prime editors (PEs) have the potential to facilitate precise genome editing in therapeutic, agricultural and research applications, their specificity has not been comprehensively evaluated. To provide a systematic assessment in plants, we first examined the mismatch tolerance of PEs in plant cells and found that the editing frequency was influenced by the number and location of mismatches in the primer binding site and spacer of the prime editing guide RNA (pegRNA). Assessing the activity of 12 pegRNAs at 179 predicted off-target sites, we detected only low frequencies of off-target edits (0.00~0.23%). Whole-genome sequencing of 29 PE-treated rice plants confirmed that PEs do not induce genome-wide pegRNA-independent off-target single-nucleotide variants or small insertions/deletions. We also show that ectopic expression of the Moloney murine leukemia virus reverse transcriptase as part of the PE does not change retrotransposon copy number or telomere structure or cause insertion of pegRNA or messenger RNA sequences into the genome. |
| 英文摘要: | Although prime editors (PEs) have the potential to facilitate precise genome editing in therapeutic, agricultural and research applications, their specificity has not been comprehensively evaluated. To provide a systematic assessment in plants, we first examined the mismatch tolerance of PEs in plant cells and found that the editing frequency was influenced by the number and location of mismatches in the primer binding site and spacer of the prime editing guide RNA (pegRNA). Assessing the activity of 12 pegRNAs at 179 predicted off-target sites, we detected only low frequencies of off-target edits (0.00~0.23%). Whole-genome sequencing of 29 PE-treated rice plants confirmed that PEs do not induce genome-wide pegRNA-independent off-target single-nucleotide variants or small insertions/deletions. We also show that ectopic expression of the Moloney murine leukemia virus reverse transcriptase as part of the PE does not change retrotransposon copy number or telomere structure or cause insertion of pegRNA or messenger RNA sequences into the genome. |
| 刊物名称: | Nature Biotechnology |
| 英文刊物名称: | Nature Biotechnology |
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| 其它备注: | Shuai Jin, Qiupeng Lin, Yingfeng Luo, Zixu Zhu, Guanwen Liu, Yunjia Li, Kunling Chen, Jin-Long Qiu and Caixia Gao. Genome-Wide Specificity of Prime Editors in Plants. Nature Biotechnology. DOI:10.1038/s41587-021-00891-x |
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