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Efficient CRISPR/Cas9-mediated Targeted Mutagenesis in Populus in the First Generation
Fan, Di1; Liu, Tingting1; Li, Chaofeng1,2; Jiao, Bo1; Li, Shuang1; Hou, Yishu1; Luo, Keming1,2
2015-07-20
发表期刊SCIENTIFIC REPORTS
卷号5
文章类型Article
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.
WOS标题词Science & Technology
关键词[WOS]CRISPR-CAS SYSTEM ; GENOME ; ARABIDOPSIS ; PLANTS ; RICE ; RECOMBINATION ; RESISTANCE ; POPLAR ; TOOL ; RNA
收录类别SCI
语种英语
WOS研究方向Science & Technology - Other Topics
WOS类目Multidisciplinary Sciences
WOS记录号WOS:000358172500002
引用统计
被引频次:116[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://210.75.249.4/handle/363003/5534
专题中国科学院西北高原生物研究所
作者单位1.Southwest Univ, Sch Life Sci, Inst Resources Bot,Minist Educ,Chongqing Key Lab, Key Lab Ecoenvironm Three Gorges Reservoir Reg, Chongqing 400715, Peoples R China
2.Chinese Acad Sci, Northwest Inst Plateau Biol, Key Lab Adaptat & Evolut Plateau Biota, Xining 810008, Peoples R China
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Fan, Di,Liu, Tingting,Li, Chaofeng,et al. Efficient CRISPR/Cas9-mediated Targeted Mutagenesis in Populus in the First Generation[J]. SCIENTIFIC REPORTS,2015,5.
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,5.
MLA Fan, Di,et al."Efficient CRISPR/Cas9-mediated Targeted Mutagenesis in Populus in the First Generation".SCIENTIFIC REPORTS 5(2015).
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