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Perfectly Matched 20-nucleotide Guide RNA Sequences Enable Robust Genome Editing Using High-fidelity SpCas9 Nucleases.
Dingbo Zhang, Huawei Zhang, Tingdong, Kunling Chen, Jinlong Qiu and Caixia Gao
Genome Biology
Abstract
High-fidelity SpCas9 variants (eSpCas9 and SpCas9-HF1) have been engineered to reduce off-target effects. We found that changes in guide RNA length induced significant reductions in the editing activities of SpCas9 variants in plant cells. Single guide RNAs harboring precise, perfectly matched 20-nucleotide guide sequences are necessary for high on-target editing activities of eSpCas9 and SpCas9-HF1. Precise 20-nucleotide guide sequences derived from tRNA–sgRNA precursors enable robust on-target editing by these variants with enhanced specificity. Our work reveals an effective way of enhancing the use of the high-fidelity SpCas9 nucleases for efficient and precise genome engineering.
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论文编号: |
DOI:10.1186/s13059-017-1325-9 |
论文题目: |
Perfectly Matched 20-nucleotide Guide RNA Sequences Enable Robust Genome Editing Using High-fidelity SpCas9 Nucleases. |
英文论文题目: |
Perfectly Matched 20-nucleotide Guide RNA Sequences Enable Robust Genome Editing Using High-fidelity SpCas9 Nucleases. |
第一作者: |
Dingbo Zhang, Huawei Zhang, Tingdong, Kunling Chen, Jinlong Qiu and Caixia Gao |
英文第一作者: |
Dingbo Zhang, Huawei Zhang, Tingdong, Kunling Chen, Jinlong Qiu and Caixia Gao |
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2017-10-12 |
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摘要: |
High-fidelity SpCas9 variants (eSpCas9 and SpCas9-HF1) have been engineered to reduce off-target effects. We found that changes in guide RNA length induced significant reductions in the editing activities of SpCas9 variants in plant cells. Single guide RNAs harboring precise, perfectly matched 20-nucleotide guide sequences are necessary for high on-target editing activities of eSpCas9 and SpCas9-HF1. Precise 20-nucleotide guide sequences derived from tRNA–sgRNA precursors enable robust on-target editing by these variants with enhanced specificity. Our work reveals an effective way of enhancing the use of the high-fidelity SpCas9 nucleases for efficient and precise genome engineering. |
英文摘要: |
High-fidelity SpCas9 variants (eSpCas9 and SpCas9-HF1) have been engineered to reduce off-target effects. We found that changes in guide RNA length induced significant reductions in the editing activities of SpCas9 variants in plant cells. Single guide RNAs harboring precise, perfectly matched 20-nucleotide guide sequences are necessary for high on-target editing activities of eSpCas9 and SpCas9-HF1. Precise 20-nucleotide guide sequences derived from tRNA–sgRNA precursors enable robust on-target editing by these variants with enhanced specificity. Our work reveals an effective way of enhancing the use of the high-fidelity SpCas9 nucleases for efficient and precise genome engineering. |
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Genome Biology |
英文刊物名称: |
Genome Biology |
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Dingbo Zhang, Huawei Zhang, Tingdong, Kunling Chen, Jinlong Qiu and Caixia Gao. Perfectly Matched 20-nucleotide Guide RNA Sequences Enable Robust Genome Editing Using High-fidelity SpCas9 Nucleases. Genome Biology. DOI:10.1186/s13059-017-1325-9 |
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