Generation of the Mutations for OsYUCCA1 in Rice Using CRISPR/Cas9 Approach
Journal Article / 2017
He Xianchang; Huang Guoqiang; Wang Daoyang; Chang Shuwei; Zhang Dabing
To investigate the role of auxin in model monocot crop rice (Oryza sativa), we constructed two vectors targeting to mutate rice auxin synthetic gene OsYUCCA1 via CRISPR/Cas9 system. The two targeted site sequences of OsYUCCA1 were synthesized and inserted into the vector called hspCas9 and then linked to the binary vector of pCAMBIA1300 . The resultant plasmids were transferred into rice through Agrobacteriummediated transformation. Three types of mutations were identified from 78 transgenic plants, including 26 homologous plants with A or T insertion respectively at 260th base on the exon and 22 heterogeneous plants with A insertion at 447th, C replaced by T at 444th and G replaced by C at 445th on the exon. Moreover, the three types of mutations of OsYUCCA1 caused a premature stop and functional loss protein in the homologous mutants. The knockout mutants of OsYUCCA1 created by CRISPR/Cas9 provides the theoretical basis for further study on the role ofauxin in rice.
|Plant Species||GE Technique
Type of Alteration
|Progress in Research