A Bright, Nontoxic, and Non-aggregating red Fluorescent Protein for Long-Term Labeling of Fine Structures in Neurons

Lin Ning; Yang Geng; Yang Geng; Matthew Lovett-Barron; Xiaoman Niu; Mengying Deng; Liang Wang; Niloufar Ataie; Alex Sens; Ho-Leung Ng; Shoudeng Chen; Karl Deisseroth; Michael Z. Lin; Michael Z. Lin; Jun Chu

doi:10.3389/fcell.2022.893468

Frontiers in Cell and Developmental Biology (Jun 2022)

A Bright, Nontoxic, and Non-aggregating red Fluorescent Protein for Long-Term Labeling of Fine Structures in Neurons

Lin Ning,
Yang Geng,
Yang Geng,
Matthew Lovett-Barron,
Xiaoman Niu,
Mengying Deng,
Liang Wang,
Niloufar Ataie,
Alex Sens,
Ho-Leung Ng,
Shoudeng Chen,
Karl Deisseroth,
Michael Z. Lin,
Michael Z. Lin,
Jun Chu

Affiliations

Lin Ning: Department of Neurobiology, Stanford University, Stanford, CA, United States
Yang Geng: Department of Neurobiology, Stanford University, Stanford, CA, United States
Yang Geng: Interdisciplinary Research Center on Biology and Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, China University of Chinese Academy of Sciences, Beijing, China
Matthew Lovett-Barron: Department of Bioengineering, Stanford University, Stanford, CA, United States
Xiaoman Niu: Interdisciplinary Research Center on Biology and Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, China University of Chinese Academy of Sciences, Beijing, China
Mengying Deng: Guangdong Provincial Key Laboratory of Biomedical Optical Imaging Technology and Center for Biomedical Optics and Molecular Imaging, and CAS Key Laboratory of Health Informatics, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China
Liang Wang: Guangdong Provincial Key Laboratory of Biomedical Optical Imaging Technology and Center for Biomedical Optics and Molecular Imaging, and CAS Key Laboratory of Health Informatics, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China
Niloufar Ataie: Department of Biochemistry and Molecular Biophysics, Kansas State University, Manhattan, KS, United States
Alex Sens: Department of Biochemistry and Molecular Biophysics, Kansas State University, Manhattan, KS, United States
Ho-Leung Ng: Department of Biochemistry and Molecular Biophysics, Kansas State University, Manhattan, KS, United States
Shoudeng Chen: Guangdong Provincial Key Laboratory of Biomedical Imaging, The Fifth Affiliated Hospital, Sun Yat-sen University, Zhuhai, China
Karl Deisseroth: Department of Bioengineering, Stanford University, Stanford, CA, United States
Michael Z. Lin: Department of Neurobiology, Stanford University, Stanford, CA, United States
Michael Z. Lin: Department of Bioengineering, Stanford University, Stanford, CA, United States
Jun Chu: Guangdong Provincial Key Laboratory of Biomedical Optical Imaging Technology and Center for Biomedical Optics and Molecular Imaging, and CAS Key Laboratory of Health Informatics, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China

DOI: https://doi.org/10.3389/fcell.2022.893468
Journal volume & issue: Vol. 10

Abstract

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Red fluorescent proteins are useful as morphological markers in neurons, often complementing green fluorescent protein-based probes of neuronal activity. However, commonly used red fluorescent proteins show aggregation and toxicity in neurons or are dim. We report the engineering of a bright red fluorescent protein, Crimson, that enables long-term morphological labeling of neurons without aggregation or toxicity. Crimson is similar to mCherry and mKate2 in fluorescence spectra but is 100 and 28% greater in molecular brightness, respectively. We used a membrane-localized Crimson-CAAX to label thin neurites, dendritic spines and filopodia, enhancing detection of these small structures compared to cytosolic markers.

Published in Frontiers in Cell and Developmental Biology

ISSN: 2296-634X (Online)
Publisher: Frontiers Media S.A.
Country of publisher: Switzerland
LCC subjects: Science: Biology (General)
Website: https://www.frontiersin.org/journals/cell-and-developmental-biology

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