基于CRISPR/ cas9的分裂荧光蛋白标记。

IF 1.4 4区生物学 Q4 DEVELOPMENTAL BIOLOGY

Zebrafish 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

Zebrafish DEVELOPMENTAL BIOLOGY-ZOOLOGY

CiteScore

3.60

自引率

5.00%

发文量

审稿时长

3 months

期刊介绍： Zebrafish is the only peer-reviewed journal dedicated to the central role of zebrafish and other aquarium species as models for the study of vertebrate development, evolution, toxicology, and human disease. Due to its prolific reproduction and the external development of the transparent embryo, the zebrafish is a prime model for genetic and developmental studies. While genetically more distant from humans, the vertebrate zebrafish nevertheless has comparable organs and tissues, such as heart, kidney, pancreas, bones, and cartilage. Zebrafish introduced the new section TechnoFish, which highlights these innovations for the general zebrafish community. TechnoFish features two types of articles: TechnoFish Previews: Important, generally useful technical advances or valuable transgenic lines TechnoFish Methods: Brief descriptions of new methods, reagents, or transgenic lines that will be of widespread use in the zebrafish community Zebrafish coverage includes: Comparative genomics and evolution Molecular/cellular mechanisms of cell growth Genetic analysis of embryogenesis and disease Toxicological and infectious disease models Models for neurological disorders and aging New methods, tools, and experimental approaches Zebrafish also includes research with other aquarium species such as medaka, Fugu, and Xiphophorus.