title: A geminivirus-based guide RNA delivery system for CRISPR/Cas9 mediated plant genome editing.
abstract: CRISPR/Cas has emerged as potent genome editing technology and has successfully been applied in many organisms, including several plant species. However, delivery of genome editing reagents remains a challenge in plants. Here, we report a virus-based guide RNA (gRNA) delivery system for CRISPR/Cas9 mediated plant genome editing (VIGE) that can be used to precisely target genome locations and cause mutations. VIGE is performed by using a modified Cabbage Leaf Curl virus (CaLCuV) vector to express gRNAs in stable transgenic plants expressing Cas9. DNA sequencing confirmed VIGE of endogenous NbPDS3 and NbIspH genes in non-inoculated leaves because CaLCuV can infect plants systemically. Moreover, VIGE of NbPDS3 and NbIspH in newly developed leaves caused photo-bleached phenotype. These results demonstrate that geminivirus-based VIGE could be a powerful tool in plant genome editing.
authors.affiliation: Center for Plant Biology, MOE Key Laboratory of Bioinformatics, School of Life Sciences, Tsinghua University, Beijing 100084, China.|State Key Laboratory of Plant Genomics, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.
authors.forename: Kangquan
authors.initials: K
authors.lastname: Yin
authors.affiliation: Center for Plant Biology, MOE Key Laboratory of Bioinformatics, School of Life Sciences, Tsinghua University, Beijing 100084, China.
authors.forename: Ting
authors.initials: T
authors.lastname: Han
authors.affiliation: Center for Plant Biology, MOE Key Laboratory of Bioinformatics, School of Life Sciences, Tsinghua University, Beijing 100084, China.
authors.forename: Guang
authors.initials: G
authors.lastname: Liu
authors.affiliation: Center for Plant Biology, MOE Key Laboratory of Bioinformatics, School of Life Sciences, Tsinghua University, Beijing 100084, China.
authors.forename: Tianyuan
authors.initials: T
authors.lastname: Chen
authors.affiliation: Center for Plant Biology, MOE Key Laboratory of Bioinformatics, School of Life Sciences, Tsinghua University, Beijing 100084, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Center for Plant Biology, MOE Key Laboratory of Bioinformatics, School of Life Sciences, Tsinghua University, Beijing 100084, China.
authors.forename: Alice Yunzi L
authors.initials: AY
authors.lastname: Yu
authors.affiliation: Center for Plant Biology, MOE Key Laboratory of Bioinformatics, School of Life Sciences, Tsinghua University, Beijing 100084, China.
authors.forename: Yule
authors.initials: Y
authors.lastname: Liu
mesh_terms: D064113:CRISPR-Cas Systems / Q000235:genetics*; D018744:DNA, Plant / Q000235:genetics* / Q000378:metabolism; D017791:Geminiviridae / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics; D018745:Genome, Plant / Q000235:genetics*; D008957:Models, Genetic; D018515:Plant Leaves / Q000235:genetics / Q000821:virology; D010944:Plants / Q000235:genetics / Q000821:virology; D030821:Plants, Genetically Modified; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000235:genetics*; D015203:Reproducibility of Results; D014026:Nicotiana / Q000235:genetics / Q000821:virology
chemical_list: D018744:DNA, Plant; D000094704:RNA, Guide, CRISPR-Cas Systems