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Plos One 10 12 : e Efficient Genome Engineering with Zinc-finger Nucleases. Molecular plant pathology, Plant Biotechnology Journal, 13 7 Clasen B. Improving cold storage and processing traits in potato through targeted gene knockout. Plant Biotechnology Journal, 14 1 Collonnier C. Plant Biotechnology Journal 15 1 DOI: Science, DOI: J and van der Zaal B.
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Precise plant breeding using new genome editing techniques: opportunities, safety and regulation in the EU. Plant Journal, 78 5 Haun W. Huang S. A proposed regulatory framework for genome-edited crops. Nature Genetics, 48 2 Hwang W. Nature Biotechnology, 31 3 DOI Plos One, 9 4 : e DOI Genome editing of the disease susceptibility gene CsLOB1 in citrus confers resistance to citrus canker.
Eukaryotic Cell, 13 11 Jiang W. Nucleic Acids Research, 41 20 : e DOI: 0. Plant Biotechnology Journal, — Science, Joung J. Journal of Experimental Botany, 66 1 Lawrenson T. Induction of targeted, heritable mutations in barley and Brassica oleracea using RNA-guided Cas9 nuclease. Genome Biology, 16 1 DOI Frontiers in Genetics, 7 DOI: Nature Biotechnology, 30 5 Lopez-Obando M.
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Plant Physiology, 2 DOI: Molecular Plant 9 7 Mahfouz M. Plant Biotechnology Journal, 12 8 Malnoy M. In eukaryotic cells, Cas9-generated DSBs are repaired by nonhomologous end joining or homologous recombination, which can be exploited to engineer insertions, deletions, and substitutions in the vicinity of the DSB. Variants of the basic targeting approach, including paired nickases Mali, Aach, et al.
Extensive biochemical and structural studies have illuminated many aspects of the molecular mechanism of Cas9. In this Chapter, we provide detailed protocols for the heterologous expression and purification of S. The assays described here can be used to validate guide RNAs and target sites for in vivo gene targeting applications or to test the in vitro efficacy of new guide RNA structures and designs.
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Moreover, the described procedures can be implemented to utilize Cas9 as a programmable restriction enzyme for DNA manipulations in vitro. Although S.
Cas9 from S. We found that expression in the Rosetta 2 strain was necessary to overcome the unfavorable codon bias in the S. Share Give access Share full text access.
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From Gene Targeting to Genome Editing: Transgenic animals applications and beyond
Abstract A set of unique sequences in bacterial genomes, responsible for protecting bacteria against bacteriophages, has recently been used for the genetic manipulation of specific points in the genome. Citing Literature. Volume 21 , Issue 4 April e