{"title":"CRISPR/Cas9-Based Split Fluorescent Protein Tagging.","authors":"Gokul Kesavan, Anja Machate, Michael Brand","doi":"10.1089/zeb.2021.0031","DOIUrl":null,"url":null,"abstract":"<p><p>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 <i>in vivo</i>.</p>","PeriodicalId":23872,"journal":{"name":"Zebrafish","volume":null,"pages":null},"PeriodicalIF":1.4000,"publicationDate":"2021-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Zebrafish","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1089/zeb.2021.0031","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2021/9/7 0:00:00","PubModel":"Epub","JCR":"Q4","JCRName":"DEVELOPMENTAL BIOLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
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.
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.
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