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Robust genome editing of CRISPR-Cas9 at NAG PAMs in rice

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Acknowledgements

This work was supported by the National Key Research and Development Program of China (2016YFD0101800), the National Natural Science Foundation of China (91635301), the Zhejiang Provincial Natural Science Foundation of China (LZ14C130003), and the Agricultural Science and Technology Innovation Program of Chinese Academy of Agricultural Sciences.

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Author notes

  1. Contributed equally to this work. X.H., X.M., J.L., and K.W. designed the studies. X.H., X.S., and X.M. performed the experiments; Q.L. conducted the bioinformatic analyses. K.W., J.L., C.G., X.H., and X.M. wrote the manuscript.

Authors and Affiliations

  1. State Key Laboratory of Plant Genomics and National Center for Plant Gene Research (Beijing), Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, 100101, China

    Xiangbing Meng, Xiaoguang Song & Jiayang Li

  2. State Key Laboratory of Rice Biology, China National Rice Research Institute, Chinese Academy of Agricultural Sciences, Hangzhou, 310006, China

    Xixun Hu, Qing Liu & Kejian Wang

  3. State Key Laboratory of Plant Cell and Chromosome Engineering, and Center for Genome Editing, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, 100101, China

    Caixia Gao

  4. University of Chinese Academy of Sciences, Beijing, 100049, China

    Jiayang Li

Authors
  1. Xiangbing Meng
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  2. Xixun Hu
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  3. Qing Liu
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  4. Xiaoguang Song
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  5. Caixia Gao
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  6. Jiayang Li
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  7. Kejian Wang
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Corresponding authors

Correspondence to Jiayang Li or Kejian Wang.

Electronic supplementary material

Figure S1

The CRISPR-Cas9 system and sequence of codon-optimized Cas9 used in this study.

Figure S2 The target locations of D1 and D2.

Figure S3 The mutation efficiency of NGG and NAG PAMs.

Figure S4 The assay of mismatch tolerance within the protospacer.

Figure S5 The target locations of NAG1 to NAG8.

Figure S6 Mutational types of rice seedlings with NAG PAMs.

Figure S7 The strategy of mixed PAMs for multiple editing.

Figure S8 The target sites of NGG and NAG in rice genome.

Table S1 The off-target effects with NAG PAM during Bph14 editing

Table S2 The location of double targets of sgRNAs

Table S3 The editing events using sgRNA with mismatches in the protospacer

Table S4 Target sites of AG1—AG8 for genome editing

Table S5 The results of off-target detection

Table S6 Primers used in the study

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Meng, X., Hu, X., Liu, Q. et al. Robust genome editing of CRISPR-Cas9 at NAG PAMs in rice. Sci. China Life Sci. 61, 122–125 (2018). https://doi.org/10.1007/s11427-017-9247-9

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  • DOI: https://doi.org/10.1007/s11427-017-9247-9

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