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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
Source PublicationSCIENTIFIC REPORTS
Volume5
SubtypeArticle
AbstractRecently, 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 HeadingsScience & Technology
WOS KeywordCRISPR-CAS SYSTEM ; GENOME ; ARABIDOPSIS ; PLANTS ; RICE ; RECOMBINATION ; RESISTANCE ; POPLAR ; TOOL ; RNA
Indexed BySCI
Language英语
WOS Research AreaScience & Technology - Other Topics
WOS SubjectMultidisciplinary Sciences
WOS IDWOS:000358172500002
Citation statistics
Cited Times:143[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://210.75.249.4/handle/363003/5534
Collection中国科学院西北高原生物研究所
Affiliation1.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
Recommended Citation
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,2015,5.
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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