基于CRISPR/ cas9的分裂荧光蛋白标记。

IF 16.4 1区化学 Q1 CHEMISTRY, MULTIDISCIPLINARY Accounts of Chemical Research Pub Date : 2021-12-01 Epub Date: 2021-09-07 DOI:10.1089/zeb.2021.0031

Gokul Kesavan, Anja Machate, Michael Brand

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引用次数: 0

摘要

基因编码的荧光标签，如绿色荧光蛋白与蛋白质融合，已经彻底改变了细胞生物学，因为它们允许在活系统中进行高分辨率的蛋白质成像。在编码序列中插入具有16个氨基酸的小片段的分裂荧光蛋白来标记蛋白质。我们成功地在斑马鱼中整合了两种明亮且快速成熟的分裂荧光蛋白，mNeon green和sfCherry2，并表明它们适合于实时成像，包括延时序列，并且具有高信噪比。此外，我们证明CRISPR/Cas9可用于在体内产生荧光标记蛋白。

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CRISPR/Cas9-Based Split Fluorescent Protein Tagging.

Genetically encoded fluorescent tags such as green fluorescent protein fused to protein have revolutionized cell biology as they permit high-resolution protein imaging in live systems. Split fluorescent proteins, with a small fragment of 16 amino acids, can be inserted in the coding sequence to label proteins. We demonstrate successful integration of two bright and fast maturing split fluorescent proteins, mNeon green and sfCherry2, in zebrafish, and show that they are suitable for live imaging, including time-lapse series, and that they have a high signal-to-noise ratio. Furthermore, we show that CRISPR/Cas9 can be used to generate fluorescently tagged proteins in vivo.

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来源期刊

Accounts of Chemical Research 化学-化学综合

CiteScore

31.40

自引率

1.10%

发文量

312

审稿时长

2 months

期刊介绍： Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance. Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.

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