Abstract:
:Homologous recombination-based gene targeting is a powerful tool for precise genome modification and has been widely used in organisms ranging from yeast to higher organisms such as Drosophila and mouse. However, gene targeting in higher plants, including the most widely used model plant Arabidopsis thaliana, remains challenging. Here we report a sequential transformation method for gene targeting in Arabidopsis. We find that parental lines expressing the bacterial endonuclease Cas9 from the egg cell- and early embryo-specific DD45 gene promoter can improve the frequency of single-guide RNA-targeted gene knock-ins and sequence replacements via homologous recombination at several endogenous sites in the Arabidopsis genome. These heritable gene targeting can be identified by regular PCR. Our approach enables routine and fine manipulation of the Arabidopsis genome.
journal_name
Nat Communjournal_title
Nature communicationsauthors
Miki D,Zhang W,Zeng W,Feng Z,Zhu JKdoi
10.1038/s41467-018-04416-0subject
Has Abstractpub_date
2018-05-17 00:00:00pages
1967issue
1issn
2041-1723pii
10.1038/s41467-018-04416-0journal_volume
9pub_type
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