METHOD FOR ACTIVATING EXPRESSION OF SILENCED GLU-1AX-NULL SUBUNIT IN WHEAT AND RELATED BIOMATERIALS

METHOD FOR ACTIVATING EXPRESSION OF SILENCED GLU-1AX-NULL SUBUNIT IN WHEAT AND RELATED BIOMATERIALS

The present invention provides a method for activating expression of a Glu-1Ax-null subunit in wheat by gene editing, the method comprising: designing a specific sgRNA for targeting a premature termination codon in a Glu-1Ax-null gene of wheat; cloning the specific sgRNA into a pair of BsaI sites of...

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Hauptverfasser: LI, Genying, LI, Jihu, GAO, Jie, ZHANG, Rongzhi, LI, Yulian, LIN, Yanxiang, LI, Wei, SONG, Guoqi, ZHANG, Shujuan
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creator LI, Genying
LI, Jihu
GAO, Jie
ZHANG, Rongzhi
LI, Yulian
LIN, Yanxiang
LI, Wei
SONG, Guoqi
ZHANG, Shujuan
description The present invention provides a method for activating expression of a Glu-1Ax-null subunit in wheat by gene editing, the method comprising: designing a specific sgRNA for targeting a premature termination codon in a Glu-1Ax-null gene of wheat; cloning the specific sgRNA into a pair of BsaI sites of a vector pEtRNA to construct a CRISPR/SpCas9 editing vector for the Glu-1Ax-null gene; and transforming the CRISPR/SpCas9 editing vector for the Glu-1Ax-null gene into wheat cell via Agrobacterium-mediated transformation and selecting a homozygous line in which the premature termination codon is disrupted. The inventive specific sgRNA is designed near the 1216-1218-site premature termination codon of the Glu-1Ax-null gene, the CRISPR/SpCas9 editing vector is transformed into wheat via Agrobacterium-mediated transformation, and the premature termination codon can be specially disrupted, thereby activating expression of silenced high-molecular-weight glutenin Glu-1Ax-null subunit in wheat.
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