title: Efficient and transgene-free genome editing in wheat through transient expression of CRISPR/Cas9 DNA or RNA.
abstract: Editing plant genomes is technically challenging in hard-to-transform plants and usually involves transgenic intermediates, which causes regulatory concerns. Here we report two simple and efficient genome-editing methods in which plants are regenerated from callus cells transiently expressing CRISPR/Cas9 introduced as DNA or RNA. This transient expression-based genome-editing system is highly efficient and specific for producing transgene-free and homozygous wheat mutants in the T0 generation. We demonstrate our protocol to edit genes in hexaploid bread wheat and tetraploid durum wheat, and show that we are able to generate mutants with no detectable transgenes. Our methods may be applicable to other plant species, thus offering the potential to accelerate basic and applied plant genome-engineering research.
authors: State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.|University of Chinese Academy of Sciences, Beijing 100049, China.
authors: Yi
authors: Y
authors: Zhang
authors: State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.|University of Chinese Academy of Sciences, Beijing 100049, China.
authors: Zhen
authors: Z
authors: Liang
authors: State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.|University of Chinese Academy of Sciences, Beijing 100049, China.
authors: Yuan
authors: Y
authors: Zong
authors: State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.|University of Chinese Academy of Sciences, Beijing 100049, China.
authors: Yanpeng
authors: Y
authors: Wang
authors: State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.
authors: Jinxing
authors: J
authors: Liu
authors: State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.
authors: Kunling
authors: K
authors: Chen
authors: State Key Laboratory of Plant Genomics, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
authors: Jin-Long
authors: JL
authors: Qiu
authors: State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.
authors: Caixia
authors: C
authors: Gao
mesh_terms: D064113:CRISPR-Cas Systems / Q000235:genetics*; D004247:DNA / Q000235:genetics; D000072669:Gene Editing / Q000331:legislation & jurisprudence / Q000379:methods*; D017343:Genes, Plant / Q000235:genetics*; D006720:Homozygote; D009154:Mutation; D030821:Plants, Genetically Modified / Q000235:genetics*; D011123:Polyploidy; D012313:RNA / Q000235:genetics; D014908:Triticum / Q000235:genetics*
chemical_list: D012313:RNA; D004247:DNA