DC pole | Wartość | Język |
dc.contributor.author | Hus, Karolina | - |
dc.contributor.author | Betekhtin, Alexander | - |
dc.contributor.author | Pinski, Artur | - |
dc.contributor.author | Rojek-Jelonek, Magdalena | - |
dc.contributor.author | Grzebelus, Ewa | - |
dc.contributor.author | Nibau, Candida | - |
dc.contributor.author | Gao, Mingjun | - |
dc.contributor.author | Jaeger, Katja E. | - |
dc.contributor.author | Jenkins, Glyn | - |
dc.contributor.author | Doonan, John H. | - |
dc.contributor.author | Hasterok, Robert | - |
dc.date.accessioned | 2020-05-21T12:52:28Z | - |
dc.date.available | 2020-05-21T12:52:28Z | - |
dc.date.issued | 2020 | - |
dc.identifier.citation | Frontiers in Plant Science, Vol. 11 (2020), art. no. 614 | pl_PL |
dc.identifier.issn | 1664-462X | - |
dc.identifier.uri | http://hdl.handle.net/20.500.12128/14180 | - |
dc.description.abstract | The CRISPR/Cas9 system enables precise genome editing and is a useful tool for
functional genomic studies. Here we report a detailed protocol for targeted genome
editing in the model grass Brachypodium distachyon and its allotetraploid relative
B. hybridum, describing gRNA design, a transient protoplast assay to test gRNA
efficiency, Agrobacterium-mediated transformation and the selection and analysis of
regenerated plants. In B. distachyon, we targeted the gene encoding phytoene
desaturase (PDS), which is a crucial enzyme in the chlorophyll biosynthesis pathway.
The albino phenotype of mutants obtained confirmed the effectiveness of the protocol
for functional gene analysis. Additionally, we targeted two genes related to cell wall
maintenance, encoding a fasciclin-like arabinogalactan protein (FLA) and a pectin
methylesterase (PME), also in B. distachyon. Two genes encoding cyclin-dependent
kinases (CDKG1 and CDKG2), which may be involved in DNA recombination were
targeted in both B. distachyon and B. hybridum. Cas9 activity induces mainly insertions
or deletions, resulting in frameshift mutations that, may lead to premature stop codons.
Because of the close phylogenetic relationship between Brachypodium species and key
temperate cereals and forage grasses, this protocol should be easily adapted to target
genes underpinning agronomically important traits. | pl_PL |
dc.language.iso | en | pl_PL |
dc.rights | Uznanie autorstwa 3.0 Polska | * |
dc.rights.uri | http://creativecommons.org/licenses/by/3.0/pl/ | * |
dc.subject | CRISPR/Cas9 system, , , , , | pl_PL |
dc.subject | targeted mutagenesis | pl_PL |
dc.subject | Brachypodium distachyon | pl_PL |
dc.subject | Brachypodium hybridum | pl_PL |
dc.subject | Agrobacterium-mediated transformation | pl_PL |
dc.subject | transient protoplast assay | pl_PL |
dc.title | A CRISPR/Cas9-Based Mutagenesis Protocol for Brachypodium distachyon and Its Allopolyploid Relative, Brachypodium hybridum | pl_PL |
dc.type | info:eu-repo/semantics/article | pl_PL |
dc.identifier.doi | 10.3389/fpls.2020.00614 | - |
Pojawia się w kolekcji: | Artykuły (WNP)
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