Targeted Mutagenesis in Hexaploid Bread Wheat Using the TALEN and CRISPR/Cas Systems
The use of sequence-specific transcription activator-like effector nucleases (TALENs) and the clustered regularly interspaced short palindromic repeats-associated system (CRISPR/Cas9) have provided powerful reverse genetic approaches to the targeted modification of genomes in numerous organisms. Both systems have been employed to generate loss-of-function alleles in bread wheat, by targeting multiple and single copies of genes. Here we present protocols for modifying the wheat genome using the two systems. The protocols include the design of TALEN and CRISPR/Cas9 target sites and their construction, evaluation of their activities in protoplasts, transformation of plants, and mutation screening.
Key wordsWheat Genome editing TALEN CRISPR/Cas9 Wheat protoplasts Targeted mutagenesis
This work was supported by grants from the National Key Research and Development Program of China (2016YFD0101804), the Chinese Academy of Sciences (KFZD-SW-107 and GJHZ1602).
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