Efficient genome editing in plants using a CRISPR/Cas system

Efficient genome editing in plants using a CRISPR/Cas system

20 August 2013 | Zhengyan Feng, Botao Zhang, Wona Ding, Xiaodong Liu, Dong-Lei Yang, Pengliang Wei, Fengqiu Cao, Shihua Zhu, Feng Zhang, Yanfei Mao, Jian-Kang Zhu
This article reports the successful application of the CRISPR/Cas system for efficient genome editing in plants. The authors engineered the *Cas9* gene and single guide RNA (sgRNA) to target specific genes in *Arabidopsis* and rice. The CRISPR/Cas system was active in creating double-strand breaks (DSBs) in *Arabidopsis* protoplasts and stably expressed in transgenic *Arabidopsis* and rice plants. The system was tested using a split yellow fluorescent protein (YFP) reporter system, showing a high gene correction rate by homologous recombination (HR). The authors then targeted endogenous loci in *Arabidopsis* and rice, including *BRASSINOSTEROID INSENSITIVE 1* (BRI1), *GIBBERELLIC ACID INSENSITIVE* (GAI), and *Rice Outermost Cell-specific gene5* (*ROC5*), *Stromal Processing Peptidase* (SPP), and *Young Seedling Albino* (*YS4*). The results showed that the CRISPR/Cas system efficiently generated mutations in these genes, leading to expected phenotypes such as dwarfism and albino leaves. The high mutation frequency and the presence of multiple mutated alleles in some transgenic plants suggest that the CRISPR/Cas system can effectively edit the plant genome, making it a promising tool for targeted gene editing in both model plants and crops.This article reports the successful application of the CRISPR/Cas system for efficient genome editing in plants. The authors engineered the *Cas9* gene and single guide RNA (sgRNA) to target specific genes in *Arabidopsis* and rice. The CRISPR/Cas system was active in creating double-strand breaks (DSBs) in *Arabidopsis* protoplasts and stably expressed in transgenic *Arabidopsis* and rice plants. The system was tested using a split yellow fluorescent protein (YFP) reporter system, showing a high gene correction rate by homologous recombination (HR). The authors then targeted endogenous loci in *Arabidopsis* and rice, including *BRASSINOSTEROID INSENSITIVE 1* (BRI1), *GIBBERELLIC ACID INSENSITIVE* (GAI), and *Rice Outermost Cell-specific gene5* (*ROC5*), *Stromal Processing Peptidase* (SPP), and *Young Seedling Albino* (*YS4*). The results showed that the CRISPR/Cas system efficiently generated mutations in these genes, leading to expected phenotypes such as dwarfism and albino leaves. The high mutation frequency and the presence of multiple mutated alleles in some transgenic plants suggest that the CRISPR/Cas system can effectively edit the plant genome, making it a promising tool for targeted gene editing in both model plants and crops.
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