Creation of gene-specific rice mutants by AvrXa23-based TALENs

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• 作者：Fu-jun WANG^a ; ^b ; ¹ ; ^{wej129@163.com} ; Chun-lian WANG^a ; ¹ ; ^{wangchunlian@caas.cn} ; Chong-ke ZHENG^a ; Teng-fei QIN^a ; Ying GAO^a ; Pi-qing LIU^b ; ^{liupq@gxu.edu.cn} ; Kai-jun ZHAO^a ; ^{zhaokaijun@caas.cn}
• 关键词：TAL effector nucleases ; AvrXa23 ; targeting ; OsERF922 ; genome editing ; rice
• 刊名：Journal of Integrative Agriculture
• 出版年：2017
• 出版时间：February 2017
• 年：2017
• 卷：16
• 期：2
• 页码：424-434
• 全文大小：1249 K
• 卷排序：16

文摘

Transcription activator-like effector (TALE) nucleases (TALENs) are increasingly used as a powerful tool for genome editing in a variety of organisms. We have previously cloned the TALE-coding gene avrXa23 from Xanthomonas oryzae pv. oryzae and developed an AvrXa23-based assembly system for designer TALEs or TALENs. Here, we exploit TALENs to induce mutagenesis of the rice ethylene response factor (ERF) transcription factor OsERF922 for testing the gene-editing efficiency of AvrXa23-based TALENs system. A pair of TALENs (T-KJ9/KJ10) was assembled and their nuclease activities were first confirmed in rice protoplast transient assay. The TALENs-expressing construct pT-KJ9/KJ10 was then used for rice transformation. We observed targeting somatic mutagenesis frequency of 15.0% in positive transgenic rice calli and obtained two mutant plants with nucleotide deletion or insertion at the designer target region. Our work demonstrates that the AvrXa23-based TALENs system can be used for site-specific genome editing in rice.