Efficient CRISPR/Cas9-mediated Targeted Mutagenesis in Populus in the First Generation
文献类型：期刊论文
| 作者 | Fan, Di ; Liu, Tingting ; Li, Chaofeng ; Jiao, Bo ; Li, Shuang ; Hou, Yishu ; Luo, Keming |
| 刊名 | SCIENTIFIC REPORTS
 |
| 出版日期 | 2015-07-20 |
| 英文摘要 | Recently, RNA-guided genome editing using the type II clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein (Cas) system has been applied to edit the plant genome in several herbaceous plant species. However, it remains unknown whether this system can be used for genome editing in woody plants. In this study, we describe the genome editing and targeted gene mutation in a woody species, Populus tomentosa Carr. via the CRISPR/Cas9 system. Four guide RNAs (gRNAs) were designed to target with distinct poplar genomic sites of the phytoene desaturase gene 8 (PtoPDS) which are followed by the protospacer-adjacent motif (PAM). After Agrobacterium-mediated transformation, obvious albino phenotype was observed in transgenic poplar plants. By analyzing the RNA-guided genome-editing events, 30 out of 59 PCR clones were homozygous mutants, 2 out of 59 were heterozygous mutants and the mutation efficiency at these target sites was estimated to be 51.7%. Our data demonstrate that the Cas9/sgRNA system can be exploited to precisely edit genomic sequence and effectively create knockout mutations in woody plants.; Recently, RNA-guided genome editing using the type II clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein (Cas) system has been applied to edit the plant genome in several herbaceous plant species. However, it remains unknown whether this system can be used for genome editing in woody plants. In this study, we describe the genome editing and targeted gene mutation in a woody species, Populus tomentosa Carr. via the CRISPR/Cas9 system. Four guide RNAs (gRNAs) were designed to target with distinct poplar genomic sites of the phytoene desaturase gene 8 (PtoPDS) which are followed by the protospacer-adjacent motif (PAM). After Agrobacterium-mediated transformation, obvious albino phenotype was observed in transgenic poplar plants. By analyzing the RNA-guided genome-editing events, 30 out of 59 PCR clones were homozygous mutants, 2 out of 59 were heterozygous mutants and the mutation efficiency at these target sites was estimated to be 51.7%. Our data demonstrate that the Cas9/sgRNA system can be exploited to precisely edit genomic sequence and effectively create knockout mutations in woody plants. |
| 源URL | [http://ir.nwipb.ac.cn/handle/363003/5534]  |
| 专题 | 西北高原生物研究所_中国科学院西北高原生物研究所 |
| 推荐引用方式 GB/T 7714 | Fan, Di,Liu, Tingting,Li, Chaofeng,et al. Efficient CRISPR/Cas9-mediated Targeted Mutagenesis in Populus in the First Generation[J]. SCIENTIFIC REPORTS,2015. |
| APA | Fan, Di.,Liu, Tingting.,Li, Chaofeng.,Jiao, Bo.,Li, Shuang.,...&Luo, Keming.(2015).Efficient CRISPR/Cas9-mediated Targeted Mutagenesis in Populus in the First Generation.SCIENTIFIC REPORTS. |
| MLA | Fan, Di,et al."Efficient CRISPR/Cas9-mediated Targeted Mutagenesis in Populus in the First Generation".SCIENTIFIC REPORTS (2015). |
入库方式： OAI收割
来源：西北高原生物研究所
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