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- Single transcript unit CRISPR 2.0 systems for robust Cas9 and Cas12a mediated plant genome editing
- xCas9 expands the scope of genome editing with reduced efficiency in rice
- Developing a highly efficient and wildly adaptive CRISPR-SaCas9 toolset for plant genome editing
- Zinc finger nuclease‐mediated precision genome editing of an endogenous gene in hexaploid bread wheat (Triticum aestivum) using a DNA repair template
- CRISPR/Cas9 editing of endogenous banana streak virus in the B genome of Musa spp. overcomes a major challenge in banana breeding
- CRISPR‐PLANT v2: An online resource for highly specific guide RNA spacers based on improved off‐target analysis
- Enhanced genome editing in rice using single transcript unit CRISPR‐LbCpf1 systems
- Efficient C-to-T base editing in plants using a fusion of nCas9 and human APOBEC3A
- Expanding the base editing scope in rice by using Cas9 variants
- Efficient Mitochondrial Genome Editing by CRISPR/Cas9
- Zinc finger nuclease‐mediated targeting of multiple transgenes to an endogenous soybean genomic locus via non‐homologous end joining
- Development of germ-line-specific CRISPR-Cas9 systems to improve the production of heritable gene modifications in Arabidopsis
- Heat-shock inducible CRISPR/Cas9 system generates heritable mutations in rice
- A CRISPR-based approach for targeted DNA demethylation
- Both CRISPR/Cas-based nucleases and nickases can be used efficiently for genome engineering in Arabidopsis thaliana
- Optimization of T-DNA architecture for Cas9-mediated mutagenesis in Arabidopsis
- Application of protoplast technology to CRISPR/Cas9 mutagenesis: from single‐cell mutation detection to mutant plant regeneration
- Comparison of efficiency and specificity of CRISPR-associated (Cas) nucleases in plants: An expanded toolkit for precision genome engineering
- Visualization analysis of CRISPR/Cas9 gene editing technology studies
- De novo domestication of wild tomato using genome editing
- Generation of inheritable and “transgene clean” targeted genome-modified rice in later generations using the CRISPR/Cas9 system
- In vivo assembly of DNA-fragments in the moss, Physcomitrella patens
- A versatile and robust Agrobacterium‐based gene stacking system generates high‐quality transgenic Arabidopsis plants
- Multiplexed Gene Editing and Protein Overexpression Using a Tobacco mosaic virus Viral Vector
- A CRISPR/Cas9 Toolbox for Multiplexed Plant Genome Editing and Transcriptional Regulation
- A CRISPR–Cpf1 system for efficient genome editing and transcriptional repression in plants
- Improved Base Editor for Efficiently Inducing Genetic Variations in Rice with CRISPR/Cas9-Guided Hyperactive hAID Mutant
- Precise A·T to G·C Base Editing in the Rice Genome
- Engineering Quantitative Trait Variation for Crop Improvement by Genome Editing
Generation of inheritable and “transgene clean” targeted genome-modified rice in later generations using the CRISPR/Cas9 system
Abstract
The CRISPR/Cas9 system is becoming an important genome editing tool for crop breeding. Although it has been demonstrated that target mutations can be transmitted to the next generation, their inheritance pattern has not yet been fully elucidated. Here, we describe the CRISPR/Cas9-mediated genome editing of four different rice genes with the help of online target-design tools. Highfrequency mutagenesis and a large percentage of putative biallelic mutations were observed in T0 generations. Nonetheless, our results also indicate that the progeny genotypes of biallelic T0 lines are frequently difficult to predict and that the transmission of mutations largely does not conform to classical genetic laws, which suggests that the mutations in T0 Transgenic rice are mainly somatic mutations. Next, we followed the inheritance pattern of T1 plants. Regardless of the presence of the CRISPR/Cas9 transgene, the mutations in T1 lines were stably transmitted to letter generations, indicating a standard germline transmission pattern. Off-target effects were also evaluated, and our results indicate that with careful target selection, off-target mutations are rare in CRISPR/ Cas9-mediated rice gene editing. Taken together, our results indicate the promising production of inheritable and “transgene clean” targeted genome-modified rice in the T1 generation using the CRISPR/Cas9 system.
Read full text here [DOi: 10.1038/srep11491]
