Efficient genome editing and gene knockout in Setaria viridis with CRISPR/Cas9 directed gene editing by the non-homologous end-joining pathway.

Autor:	Basso MF; National Center for Agroenergy Research (CNPAE), Brazilian Agricultural Research Corporation (EMBRAPA), Brasília, DF, 70770-901, Brazil.; BIOMOL/BIOTEC Laboratory, Mato Grosso Cotton Institute (IMAmt), Rondonópolis, MT, 78740-970, Brazil., Duarte KE; National Center for Agroenergy Research (CNPAE), Brazilian Agricultural Research Corporation (EMBRAPA), Brasília, DF, 70770-901, Brazil.; Center for Natural and Human Sciences, Federal University of ABC (UFABC), São Bernardo do Campo, São Paulo, 09606-045, Brazil., Santiago TR; National Center for Agroenergy Research (CNPAE), Brazilian Agricultural Research Corporation (EMBRAPA), Brasília, DF, 70770-901, Brazil.; Departament of Phytopathology, Federal University of Brasília, Brasília (UNB), Distrito Federal, 70910-900, Brazil., de Souza WR; Center for Natural and Human Sciences, Federal University of ABC (UFABC), São Bernardo do Campo, São Paulo, 09606-045, Brazil., Garcia BO; National Center for Agroenergy Research (CNPAE), Brazilian Agricultural Research Corporation (EMBRAPA), Brasília, DF, 70770-901, Brazil., da Cunha BDB; National Center for Agroenergy Research (CNPAE), Brazilian Agricultural Research Corporation (EMBRAPA), Brasília, DF, 70770-901, Brazil., Kobayashi AK; National Center for Agroenergy Research (CNPAE), Brazilian Agricultural Research Corporation (EMBRAPA), Brasília, DF, 70770-901, Brazil., Molinari HBC; National Center for Agroenergy Research (CNPAE), Brazilian Agricultural Research Corporation (EMBRAPA), Brasília, DF, 70770-901, Brazil.
Jazyk:	angličtina
Zdroj:	Plant biotechnology (Tokyo, Japan) [Plant Biotechnol (Tokyo)] 2021 Jun 25; Vol. 38 (2), pp. 227-238.
DOI:	10.5511/plantbiotechnology.21.0407a
Abstrakt:	The CRISPR/Cas9 system has been used for genome editing in several organisms, including higher plants. This system induces site-specific mutations in the genome based on the nucleotide sequence of engineered guide RNAs. The complex genomes of C4 grasses makes genome editing a challenge in key grass crops like maize ( Zea mays ), sorghum ( Sorghum bicolor ), Brachiaria spp., switchgrass ( Panicum virgatum ), and sugarcane ( Saccharum spp.). Setaria viridis is a diploid C4 grass widely used as a model for these C4 crop plants. Here, an optimized CRISPR/Cas9 binary vector that exploits the non-homologous end joining (NHEJ) system was used to knockout a green fluorescent protein ( gfp ) transgene in S. viridis accession A10.1. Transformation of embryogenic callus by A. tumefaciens generated ten glufosinate-ammonium resistant transgenic events. In the T0 generation, 60% of the events were biallelic mutants in the gfp transgene with no detectable accumulation of GFP protein and without insertions or deletions in predicted off-target sites. The gfp mutations generated by CRISPR/Cas9 were stable and displayed Mendelian segregation in the T1 generation. Altogether, the system described here is a highly efficient genome editing system for S. viridis , an important model plant for functional genomics studies in C4 grasses. Also, this system is a potential tool for improvement of agronomic traits in C4 crop plants with complex genomes. (© 2021 Japanese Society for Plant Biotechnology.)
Databáze:	MEDLINE
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