CRISPR/Cas9-mediated genome editing and gene replacement in plants: Transitioning from lab to field

Schaeffer, Scott; Nakata, Paul A.

doi:10.1016/j.plantsci.2015.09.011

Cited by 147 publications

(83 citation statements)

References 111 publications

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“…New sequencing approaches combined with an increase in identification of candidate genes and more precise techniques of genetic engineering will have a big impact on plant breeding, especially since the new methods like CRISPR-Cas9 are much easier and cheaper in their application than former methods [48][49][50][51]. The potential advantages of these new techniques have been widely published.…”

Section: Continuous Developments and The Need For Clear Evaluation Crmentioning

confidence: 99%

Concepts and Strategies of Organic Plant Breeding in Light of Novel Breeding Techniques

2016

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Abstract:In this paper, we describe the development of a set of guiding principles for the evaluation of breeding techniques by the organic sector over time. The worldwide standards of organic agriculture (OA) do not allow genetic engineering (GE) or any products derived from genetic engineering. The standards in OA are an expression of the underlying principles of health, ecology, fairness and care. The derived norms are process and not product oriented. As breeding is considered part of the process in agriculture, GE is not a neutral tool for the organic sector. The incompatibility between OA and GE is analyzed, including the "novel breeding techniques". Instead, alternative breeding approaches are pursued based on the norms and values of organic agriculture not only on the technical level but also on the social and organizational level by including other value chain players and consumers. The status and future perspectives of the alternative directions for organic breeding are described and discussed.

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Section: Continuous Developments and The Need For Clear Evaluation Crmentioning

confidence: 99%

Concepts and Strategies of Organic Plant Breeding in Light of Novel Breeding Techniques

2016

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“…The type II CRISPR-Cas9 system has been extensively developed as a powerful genome-editing tool because of its high on-target efficiency in numerous prokaryotes and eukaryotes, including (but not limited to) E. coli [16], Streptomyces spp. [7], Clostridium cellulolyticum [43], Lactobacillus reuteri [30], Saccharomyces cerevisiae [11], Bombyx mori [41], Filamentous fungi [23,28], Drosophila [46], Candida albicans [40], higher plants [32], and multiple human cell lines [8,19,25]. The CRISPR-associated protein (Cas9) endonuclease is guided by a mature CRISPR RNA (crRNA) and a trans-activating crRNA (tracrRNA) towards a target DNA sequence (known as a protospacer).…”

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confidence: 99%

Multiplex gene editing of the Yarrowia lipolytica genome using the CRISPR-Cas9 system

Gao

Tong

Wen

et al. 2016

Journal of Industrial Microbiology and Biotechnology

161

143

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Yarrowia lipolytica is categorized as a generally recognized as safe (GRAS) organism and is a heavily documented, unconventional yeast that has been widely incorporated into multiple industrial fields to produce valuable biochemicals. This study describes the construction of a CRISPR-Cas9 system for genome editing in Y. lipolytica using a single plasmid (pCAS1yl or pCAS2yl) to transport Cas9 and relevant guide RNA expression cassettes, with or without donor DNA, to target genes. Two Cas9 target genes, TRP1 and PEX10, were repaired by non-homologous end-joining (NHEJ) or homologous recombination, with maximal efficiencies in Y. lipolytica of 85.6 % for the wild-type strain and 94.1 % for the ku70/ku80 double-deficient strain, within 4 days. Simultaneous double and triple multigene editing was achieved with pCAS1yl by NHEJ, with efficiencies of 36.7 or 19.3 %, respectively, and the pCASyl system was successfully expanded to different Y. lipolytica breeding strains. This timesaving method will enable and improve synthetic biology, metabolic engineering and functional genomic studies of Y. lipolytica.

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“…Of the three types of CRISPR/Cas systems, type II has been most intensively studied and developed for genome engineering techniques in eukaryotes, including plants3456789.…”

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confidence: 99%

Highly specific targeted mutagenesis in plants using Staphylococcus aureus Cas9

Kaya

Mikami

Endo

et al. 2016

Sci Rep

113

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The CRISPR/Cas9 system is an efficient and convenient tool for genome editing in plants. Cas9 nuclease derived from Streptococcus pyogenes (Sp) is commonly used in this system. Recently, Staphylococcus aureus Cas9 (SaCas9)-mediated genome editing was reported in human cells and Arabidopsis. Because SaCas9 (1053 a.a.) is smaller than SpCas9 (1368 a.a.), SaCas9 could have substantial advantages for delivering and expressing Cas9 protein, especially using virus vectors. Since the protospacer adjacent motif (PAM) sequence of SaCas9 (5′-NNGRRT-3′) differs from that of SpCas9 (5′-NGG-3′), the use of this alternative Cas9 nuclease could expand the selectivity at potential cleavage target sites of the CRISPR/Cas9 system. Here we show that SaCas9 can mutagenize target sequences in tobacco and rice with efficiencies similar to those of SpCas9. We also analyzed the base preference for ‘T’ at the 6th position of the SaCas9 PAM. Targeted mutagenesis efficiencies in target sequences with non-canonical PAMs (5′-NNGRRV-3′) were much lower than those with a canonical PAM (5′-NNGRRT-3′). The length of target sequence recognized by SaCas9 is one or two nucleotides longer than that recognized by SpCas9. Taken together, our results demonstrate that SaCas9 has higher sequence recognition capacity than SpCas9 and is useful for reducing off-target mutations in crop.

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CRISPR/Cas9-mediated genome editing and gene replacement in plants: Transitioning from lab to field

Cited by 147 publications

References 111 publications

Concepts and Strategies of Organic Plant Breeding in Light of Novel Breeding Techniques

Concepts and Strategies of Organic Plant Breeding in Light of Novel Breeding Techniques

Multiplex gene editing of the Yarrowia lipolytica genome using the CRISPR-Cas9 system

Highly specific targeted mutagenesis in plants using Staphylococcus aureus Cas9

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