Chapter title |
Design and Assembly of CRISPR/Cas9 Reagents for Gene Knockout, Targeted Insertion, and Replacement in Wheat
|
---|---|
Chapter number | 12 |
Book title |
Wheat Biotechnology
|
Published in |
Methods in molecular biology, January 2017
|
DOI | 10.1007/978-1-4939-7337-8_12 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7335-4, 978-1-4939-7337-8
|
Authors |
Tomáš Čermák, Shaun J. Curtin |
Abstract |
Advances in cereal transformation along with the completion of the wheat genome sequence assembly have increased the demand for tools that perform targeted and specific modifications in this crop plant. This protocol demonstrates the construction of reagents using a comprehensive genome engineering kit to create single and multiple gene "knockouts," site-specific chromosome deletions and gene replacement or "knockins" including the use of geminivirus replicons (GVRs). The reagents allow for both easy construction of simple genome engineering vectors, and "mix and match" swapping of components such as the Cas9, guide RNA and donor template cassettes for gene targeting. In addition, a web-based tool greatly streamlines vector selection, primer design, and vector construction. |
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Geographical breakdown
Country | Count | As % |
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Unknown | 4 | 100% |
Demographic breakdown
Type | Count | As % |
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Members of the public | 4 | 100% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 26 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
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Researcher | 10 | 38% |
Student > Ph. D. Student | 4 | 15% |
Student > Bachelor | 3 | 12% |
Other | 2 | 8% |
Professor | 1 | 4% |
Other | 0 | 0% |
Unknown | 6 | 23% |
Readers by discipline | Count | As % |
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Biochemistry, Genetics and Molecular Biology | 9 | 35% |
Agricultural and Biological Sciences | 8 | 31% |
Immunology and Microbiology | 1 | 4% |
Engineering | 1 | 4% |
Unknown | 7 | 27% |