摘要
Transcription activator-like effector （TALE） nucleases （TALENs） are increasingly used as a powerful tool for genome edit- ing 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/KJ 10） 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.
Transcription activator-like effector （TALE） nucleases （TALENs） are increasingly used as a powerful tool for genome edit- ing 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/KJ 10） 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.
基金
supported by the grant from the Major Science and Technology Project to Create New Crop Cultivars Using Gene Transfer Technology of China (2014ZX0801001B)
the National Natural Science Foundation of China (31171812)
