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| 003 | MX-MdCICY | ||
| 005 | 20250625160158.0 | ||
| 040 | _cCICY | ||
| 090 | _aB-16780 | ||
| 245 | 1 | 0 | _aOn Improving CRISPR for Editing Plant Genes: Ribozyme-Mediated Guide RNA Production and Fluorescence-Based Technology for Isolating Transgene-Free Mutants Generated by CRISPR |
| 490 | 0 | _vProgress in Molecular Biology and Translational Science, 149, p.151-166, 2017 | |
| 520 | 3 | _aCRISPR/Cas9-mediated genome editing technology has been used to successfully edit numerous genes in various organisms including plants. There are still two major challenges in using CRISPR/Cas9 technology for gene editing in plants. First, there are very limited choices of promoters that are suitable for in vivo production of single-guide RNAs (sgRNAs), which is complementary to the target sequence and which guides Cas9 to generate double-strand breaks at the target site. It is especially difficult to produce sgRNA molecules with temporal and spatial precision. Second, there is a lack of efficient methods for identifying plants that (1)contain heritable and stable mutations generated by CRISPR/Cas9, and (2)no longer harbor the CRISPR/Cas9 construct and other transgenes. In this chapter, we describe the development of a ribozyme-based strategy that enables the production of sgRNA molecules from any chosen promoter. More importantly, the ribozyme-based technology makes it feasible to produce sgRNAs with temporal and spatial precision, greatly expanding the scope and applications of CRISPR/Cas9 technology. We also developed a fluorescence-based technology that allows us to efficiently and reliably isolate Cas9-free stable Arabidopsis mutants. Thus, we provide effective protocols to overcome two important obstacles in using CRISPR/Cas9 for editing genes in plants. | |
| 650 | 1 | 4 | _aARABIDOPSIS |
| 650 | 1 | 4 | _aCAS9 |
| 650 | 1 | 4 | _aCRISPR |
| 650 | 1 | 4 | _aMCHERRY |
| 650 | 1 | 4 | _aRGR |
| 650 | 1 | 4 | _aRIBOZYME |
| 700 | 1 | 2 | _aHe, Y. |
| 700 | 1 | 2 | _aWang, R. |
| 700 | 1 | 2 | _aDai, X. |
| 700 | 1 | 2 | _aZhao, Y. |
| 856 | 4 | 0 |
_uhttps://drive.google.com/file/d/1rpAuNa34agjO6ut02kQcxYMIN_ejwXZ4/view?usp=drivesdk _zPara ver el documento ingresa a Google con tu cuenta: @cicy.edu.mx |
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