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

Autor: Fu-jun WANG, Chun-lian WANG, Chong-ke ZHENG, Teng-fei QIN, Ying GAO, Pi-qing LIU, Kai-jun ZHAO
Jazyk: angličtina
Rok vydání: 2017
Předmět:
Zdroj: Journal of Integrative Agriculture, Vol 16, Iss 2, Pp 424-434 (2017)
Druh dokumentu: article
ISSN: 2095-3119
DOI: 10.1016/S2095-3119(16)61411-9
Popis: 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.
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