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优化用于硅藻的CRISPR/Cas9

Optimizing CRISPR/Cas9 for the Diatom .

作者信息

Stukenberg Daniel, Zauner Stefan, Dell'Aquila Gianluca, Maier Uwe G

机构信息

Department for Cell Biology, Philipps-Universität Marburg, Marburg, Germany.

LOEWE-Zentrum für Synthetische Mikrobiologie (SYNMIKRO), Philipps-Universität Marburg, Marburg, Germany.

出版信息

Front Plant Sci. 2018 Jun 6;9:740. doi: 10.3389/fpls.2018.00740. eCollection 2018.

Abstract

CRISPR/Cas9 is a powerful tool for genome editing. We constructed an easy-to-handle expression vector for application in the model organism and tested its capabilities in order to apply CRISPR/Cas9 technology for our purpose. In our experiments, we targeted two different genes, screened for mutations and analyzed mutated diatoms in a three-step process. In the end, we identified cells, showing either monoallelic or homo-biallelic targeted mutations. Thus, we confirm that application of the CRISPR/Cas9 system for is very promising, although, as discussed, overlooked pitfalls have to be considered.

摘要

CRISPR/Cas9是一种用于基因组编辑的强大工具。我们构建了一种易于操作的表达载体,用于在模式生物中应用,并测试了其功能,以便将CRISPR/Cas9技术应用于我们的目的。在我们的实验中,我们针对两个不同的基因,通过一个三步过程筛选突变并分析突变的硅藻。最后,我们鉴定出显示单等位基因或双等位基因靶向突变的细胞。因此,我们证实,尽管如所讨论的那样,必须考虑到被忽视的陷阱,但将CRISPR/Cas9系统应用于[此处原文缺失具体应用内容]是非常有前景的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/22de/5998643/572ffc85d3d1/fpls-09-00740-g001.jpg

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