Generation of inheritable and "transgene clean" targeted genome-modified rice in later generations using the CRISPR/Cas9 system

The CRISPR/Cas9 system is becoming an important genome editing tool for crop breeding. Although it has been demonstrated that target mutations can be transmitted to the next generation, their inheritance pattern has not yet been fully elucidated. Here, we describe the CRISPR/Cas9-mediated genome edi...

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Bibliographic Details
Published in:Scientific reports 2015-06, Vol.5 (1), p.11491, Article 11491
Main Authors: Xu, Rong-Fang, Li, Hao, Qin, Rui-Ying, Li, Juan, Qiu, Chun-Hong, Yang, Ya-Chun, Ma, Hui, Li, Li, Wei, Peng-Cheng, Yang, Jian-Bo
Format: Article
Language:English
Subjects:
CRISPR
CRISPR-Cas Systems
Gene Targeting
Genes, Plant
Genome editing
Genome, Plant
Genomes
Genomic Instability
Genotypes
Heredity
Inheritance Patterns
Mutagenesis
Mutation
Oryza - genetics
Plant breeding
Plants, Genetically Modified
Rice
RNA Editing
Transgenes
Transgenic plants
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Summary:The CRISPR/Cas9 system is becoming an important genome editing tool for crop breeding. Although it has been demonstrated that target mutations can be transmitted to the next generation, their inheritance pattern has not yet been fully elucidated. Here, we describe the CRISPR/Cas9-mediated genome editing of four different rice genes with the help of online target-design tools. High-frequency mutagenesis and a large percentage of putative biallelic mutations were observed in T0 generations. Nonetheless, our results also indicate that the progeny genotypes of biallelic T0 lines are frequently difficult to predict and that the transmission of mutations largely does not conform to classical genetic laws, which suggests that the mutations in T0 transgenic rice are mainly somatic mutations. Next, we followed the inheritance pattern of T1 plants. Regardless of the presence of the CRISPR/Cas9 transgene, the mutations in T1 lines were stably transmitted to later generations, indicating a standard germline transmission pattern. Off-target effects were also evaluated, and our results indicate that with careful target selection, off-target mutations are rare in CRISPR/Cas9-mediated rice gene editing. Taken together, our results indicate the promising production of inheritable and "transgene clean" targeted genome-modified rice in the T1 generation using the CRISPR/Cas9 system.
ISSN:2045-2322
2045-2322
DOI:10.1038/srep11491