| 000 | 02689nab a22004217a 4500 | ||
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| 999 |
_c58263 _d58255 |
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| 001 | 58263 | ||
| 003 | MX-TxCIM | ||
| 005 | 20191010225020.0 | ||
| 008 | 161010s2016 -uk|||p|op||| 00| 00eng d | ||
| 024 | 8 | _ahttps://doi.org/10.1038/srep23890 | |
| 040 | _aMX-TxCIM | ||
| 041 | _aeng | ||
| 100 | 0 |
_94198 _aYongping Zhao |
|
| 245 | 1 | 0 |
_aAn alternative strategy for targeted gene replacement in plants using a dual-sgRNA/Cas9 design _h[Electronic Resource] |
| 260 |
_aLondon : _bNature Publishing Group, _c2016. |
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| 500 | _aPeer review | ||
| 500 | _aOpen Access | ||
| 520 | _aPrecision DNA/gene replacement is a promising genome-editing tool that is highly desirable for molecular engineering and breeding by design. Although the CRISPR/Cas9 system works well as a tool for gene knockout in plants, gene replacement has rarely been reported. Towards this end, we first designed a combinatory dual-sgRNA/Cas9 vector (construct #1) that successfully deleted miRNA gene regions (MIR169a and MIR827a). The deletions were confirmed by PCR and subsequent sequencing, yielding deletion efficiencies of 20% and 24% on MIR169a and MIR827a loci, respectively. We designed a second structure (construct #2) that contains sites homologous to Arabidopsis TERMINAL FLOWER 1 (TFL1) for homology-directed repair (HDR) with regions corresponding to the two sgRNAs on the modified construct #1. The two constructs were co-transformed into Arabidopsis plants to provide both targeted deletion and donor repair for targeted gene replacement by HDR. Four of 500 stably transformed T0 transgenic plants (0.8%) contained replaced fragments. The presence of the expected recombination sites was further confirmed by sequencing. Therefore, we successfully established a gene deletion/replacement system in stably transformed plants that can potentially be utilized to introduce genes of interest for targeted crop improvement. | ||
| 526 |
_aMCRP _bFP2 _bFP3 |
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| 546 | _aText in English | ||
| 591 | _bCIMMYT Informa: 1989 (April 20, 2017) | ||
| 650 | 7 |
_93563 _aGenes _2AGROVOC |
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| 650 | 7 |
_94199 _aPlants _2AGROVOC |
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| 650 | 7 |
_98831 _aGenetic engineering _gAGROVOC |
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| 650 | 7 |
_93444 _aMolecular genetics _2AGROVOC |
|
| 700 | 0 |
_94200 _aCongsheng Zhang |
|
| 700 | 0 |
_94201 _aWenwen Liu |
|
| 700 | 0 |
_94202 _aWei Gao |
|
| 700 | 0 |
_94203 _aChanglin Liu |
|
| 700 | 0 |
_94204 _aGaoyuan Song |
|
| 700 | 0 |
_92733 _aWen-Xue Li |
|
| 700 | 0 |
_94205 _aLong Mao |
|
| 700 | 0 |
_94206 _aBeijiu Chen |
|
| 700 | 1 |
_9857 _aYunbi Xu _gGlobal Maize Program _8INT2735 |
|
| 700 | 0 |
_94207 _aXinhai Li |
|
| 700 | 0 |
_92730 _aChuanxiao Xie |
|
| 773 | 0 |
_wa58025 _x2045-2322 _dLondon : Nature Publishing Group, 2011- _tNature Scientific reports _gv. 6, no. 23890 |
|
| 856 | 4 |
_uhttp://hdl.handle.net/10883/18311 _yOpen Access through DSpace |
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| 942 |
_2ddc _cJA _n0 |
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