A versatile toolkit for CRISPR-Cas13-based RNA manipulation in Drosophila.

A versatile toolkit for CRISPR-Cas13-based RNA manipulation in Drosophila.
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一个基于CRISPR-Cas13的果蝇RNA操作通用工具包。

DOI:
10.1186/s13059-020-02193-y
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发表时间:
2020-11-17
期刊:
影响因子:
12.3
通讯作者:
King-Jones K
King-Jones K
中科院分区:
生物学1区
文献类型:
--
作者:
Huynh N;Depner N;Larson R;King-Jones K

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CRISPR技术的进步极大地提高了我们操纵核酸的能力,最近发现的RNA靶向核酸内切酶Cas 13甚至增加了更多的功能。在这里,我们表明Cas 13在果蝇体内和体外都有效地起作用。我们测试了44种不同的Cas 13变体,以鉴定具有最佳整体性能的酶,并表明Cas 13可以高效和特异性地靶向体内内源性果蝇转录物。我们还开发了Cas 13应用程序来编辑mRNA和靶向线粒体转录本。我们的载体集合代表了在转录后水平操纵基因表达的多功能工具集合。在线版本包含补充材料,可通过10.1186/s13059-020-02193-y获得。
Advances in CRISPR technology have immensely improved our ability to manipulate nucleic acids, and the recent discovery of the RNA-targeting endonuclease Cas13 adds even further functionality. Here, we show that Cas13 works efficiently in Drosophila, both ex vivo and in vivo. We test 44 different Cas13 variants to identify enzymes with the best overall performance and show that Cas13 could target endogenous Drosophila transcripts in vivo with high efficiency and specificity. We also develop Cas13 applications to edit mRNAs and target mitochondrial transcripts. Our vector collection represents a versatile tool collection to manipulate gene expression at the post-transcriptional level. The online version contains supplementary material available at 10.1186/s13059-020-02193-y.
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