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| Obtainment and Characterization of Rice (Oryza sativa) OsDUF1475 Mutants Based on CRISPR/Cas9 Technique |
| KONG Xiao-Cong, PI Rui-Xue, SHI Yu-Lu, WANG Quan, JIN Ya-Jun, LIANG Shan-Shan, ZHANG Si-Ju, LUAN Wei-Jiang* |
| Tianjin Key Laboratory of Animal and Plant Resistance/College of Life Sciences, Tianjin Normal University, Tianjin 300387, China |
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Abstract Domains of unknown function protein families (DUFs) are a large set of uncharacterized protein families, which play an important role in biological activities. To investigate biological function of the rice (Oryza sativa) DUF gene OsDUF1475, the CRISPR/Cas9 gene editing technique was employed. Two target sites on the first exon of the OsDUF1475 gene were selected and ligased into CRISPR/Cas9 vector to produce 2 recombinant vectors. Then, the 2 recombined plasmids were introduced into rice cultivar Nipponbare by Agrobacterium-mediated transformation method. Totally 65 T0 transgenic plants were obtained. Sequencing analysis of the target sites showed that there were 3 types of mutations in the transgenic plants, including single base insertions, short fragment deletions (<10 bp) and long fragment deletions (>10 bp). Analysis of the amino acid sequence showed that the frame shift of OsDUF1475 ORF were produced which resulted in truncated protein in editing mutants. The results indicated that the OsDUF1475 gene was edited successfully by CRISPR/Cas9 technique. And T1 generation homozygous mutant plants obtained by self-crossing showed that these mutants were genetically stable. Taken together, the mutants obtained in present study are suitable materials for OsDUF1475 function study.
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Received: 31 August 2018
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Corresponding Authors:
* lwjzsq@163.com
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