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1.
Nat Plants ; 4(9): 730, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-30139955

RESUMO

In Supplementary Fig. 1b originally published with this Brief Communication, the DNA sequence of nickase Cas9 was incorrect; this has now been amended.

2.
Nat Plants ; 4(7): 427-431, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-29867128

RESUMO

The recent development of adenine base editors (ABEs) has enabled efficient and precise A-to-G base conversions in higher eukaryotic cells. Here, we show that plant-compatible ABE systems can be successfully applied to protoplasts of Arabidopsis thaliana and Brassica napus through transient transfection, and to individual plants through Agrobacterium-mediated transformation to obtain organisms with desired phenotypes. Targeted, precise A-to-G substitutions generated a single amino acid change in the FT protein or mis-splicing of the PDS3 RNA transcript, and we could thereby obtain transgenic plants with late-flowering and albino phenotypes, respectively. Our results provide 'proof of concept' for in planta ABE applications that can lead to induced neo-functionalization or altered mRNA splicing, opening up new avenues for plant genome engineering and biotechnology.


Assuntos
Adenina , Edição de Genes/métodos , Engenharia Genética/métodos , Genoma de Planta/genética , Arabidopsis/genética , Brassica napus/genética , Proteína 9 Associada à CRISPR , Sistemas CRISPR-Cas , Plantas Geneticamente Modificadas , Protoplastos
3.
Nat Biotechnol ; 33(11): 1162-4, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26479191

RESUMO

Editing plant genomes without introducing foreign DNA into cells may alleviate regulatory concerns related to genetically modified plants. We transfected preassembled complexes of purified Cas9 protein and guide RNA into plant protoplasts of Arabidopsis thaliana, tobacco, lettuce and rice and achieved targeted mutagenesis in regenerated plants at frequencies of up to 46%. The targeted sites contained germline-transmissible small insertions or deletions that are indistinguishable from naturally occurring genetic variation.


Assuntos
Sistemas CRISPR-Cas/genética , Engenharia Genética/métodos , Genoma de Planta/genética , Plantas Geneticamente Modificadas/genética , Ribonucleoproteínas/genética , Arabidopsis/genética , Arabidopsis/fisiologia , Sequência de Bases , Técnicas de Inativação de Genes , Dados de Sequência Molecular , Oryza/genética , Oryza/fisiologia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/fisiologia , Protoplastos , Ribonucleoproteínas/metabolismo , Nicotiana/genética , Nicotiana/fisiologia
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