Biotagging of Specific Cell Populations in Zebrafish Reveals Gene Regulatory Logic Encoded in the Nuclear Transcriptome

Summary: Interrogation of gene regulatory circuits in complex organisms requires precise tools for the selection of individual cell types and robust methods for biochemical profiling of target proteins. We have developed a versatile, tissue-specific binary in vivo biotinylation system in zebrafish t...

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Main Authors: Le A. Trinh, Vanessa Chong-Morrison, Daria Gavriouchkina, Tatiana Hochgreb-Hägele, Upeka Senanayake, Scott E. Fraser, Tatjana Sauka-Spengler
Format: Article
Language:English
Published: Elsevier 2017-04-01
Series:Cell Reports
Online Access:http://www.sciencedirect.com/science/article/pii/S2211124717303911
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author Le A. Trinh
Vanessa Chong-Morrison
Daria Gavriouchkina
Tatiana Hochgreb-Hägele
Upeka Senanayake
Scott E. Fraser
Tatjana Sauka-Spengler
author_facet Le A. Trinh
Vanessa Chong-Morrison
Daria Gavriouchkina
Tatiana Hochgreb-Hägele
Upeka Senanayake
Scott E. Fraser
Tatjana Sauka-Spengler
author_sort Le A. Trinh
collection DOAJ
description Summary: Interrogation of gene regulatory circuits in complex organisms requires precise tools for the selection of individual cell types and robust methods for biochemical profiling of target proteins. We have developed a versatile, tissue-specific binary in vivo biotinylation system in zebrafish termed biotagging that uses genetically encoded components to biotinylate target proteins, enabling in-depth genome-wide analyses of their molecular interactions. Using tissue-specific drivers and cell-compartment-specific effector lines, we demonstrate the specificity of the biotagging toolkit at the biochemical, cellular, and transcriptional levels. We use biotagging to characterize the in vivo transcriptional landscape of migratory neural crest and myocardial cells in different cellular compartments (ribosomes and nucleus). These analyses reveal a comprehensive network of coding and non-coding RNAs and cis-regulatory modules, demonstrating that tissue-specific identity is embedded in the nuclear transcriptomes. By eliminating background inherent to complex embryonic environments, biotagging allows analyses of molecular interactions at high resolution. : A genetically encoded in vivo biotinylation system in zebrafish developed by Trinh et al. reveals cell-type- and subcellular-compartment-specific coding and non-coding RNAs in developing cardiomyocytes and neural crest cells. Characterization of non-coding RNAs in neural crest reveals bidirectionally transcribed cis-regulatory elements that define a specific gene regulatory signature. Keywords: in vivo biotinylation, nuclear transcriptome, neural crest, myocardium, enhancers, cis-regulation, bi-directional transcription
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spelling doaj.art-6f8fee874fdc4d49b181a641e77d55ef2022-12-22T01:41:50ZengElsevierCell Reports2211-12472017-04-01192425440Biotagging of Specific Cell Populations in Zebrafish Reveals Gene Regulatory Logic Encoded in the Nuclear TranscriptomeLe A. Trinh0Vanessa Chong-Morrison1Daria Gavriouchkina2Tatiana Hochgreb-Hägele3Upeka Senanayake4Scott E. Fraser5Tatjana Sauka-Spengler6Molecular and Computational Biology, University of Southern California, Los Angeles, CA 90089, USARadcliffe Department of Medicine, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford OX3 9DS, UKRadcliffe Department of Medicine, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford OX3 9DS, UKRadcliffe Department of Medicine, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford OX3 9DS, UK; Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA 91125, USARadcliffe Department of Medicine, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford OX3 9DS, UKMolecular and Computational Biology, University of Southern California, Los Angeles, CA 90089, USARadcliffe Department of Medicine, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford OX3 9DS, UK; Corresponding authorSummary: Interrogation of gene regulatory circuits in complex organisms requires precise tools for the selection of individual cell types and robust methods for biochemical profiling of target proteins. We have developed a versatile, tissue-specific binary in vivo biotinylation system in zebrafish termed biotagging that uses genetically encoded components to biotinylate target proteins, enabling in-depth genome-wide analyses of their molecular interactions. Using tissue-specific drivers and cell-compartment-specific effector lines, we demonstrate the specificity of the biotagging toolkit at the biochemical, cellular, and transcriptional levels. We use biotagging to characterize the in vivo transcriptional landscape of migratory neural crest and myocardial cells in different cellular compartments (ribosomes and nucleus). These analyses reveal a comprehensive network of coding and non-coding RNAs and cis-regulatory modules, demonstrating that tissue-specific identity is embedded in the nuclear transcriptomes. By eliminating background inherent to complex embryonic environments, biotagging allows analyses of molecular interactions at high resolution. : A genetically encoded in vivo biotinylation system in zebrafish developed by Trinh et al. reveals cell-type- and subcellular-compartment-specific coding and non-coding RNAs in developing cardiomyocytes and neural crest cells. Characterization of non-coding RNAs in neural crest reveals bidirectionally transcribed cis-regulatory elements that define a specific gene regulatory signature. Keywords: in vivo biotinylation, nuclear transcriptome, neural crest, myocardium, enhancers, cis-regulation, bi-directional transcriptionhttp://www.sciencedirect.com/science/article/pii/S2211124717303911
spellingShingle Le A. Trinh
Vanessa Chong-Morrison
Daria Gavriouchkina
Tatiana Hochgreb-Hägele
Upeka Senanayake
Scott E. Fraser
Tatjana Sauka-Spengler
Biotagging of Specific Cell Populations in Zebrafish Reveals Gene Regulatory Logic Encoded in the Nuclear Transcriptome
Cell Reports
title Biotagging of Specific Cell Populations in Zebrafish Reveals Gene Regulatory Logic Encoded in the Nuclear Transcriptome
title_full Biotagging of Specific Cell Populations in Zebrafish Reveals Gene Regulatory Logic Encoded in the Nuclear Transcriptome
title_fullStr Biotagging of Specific Cell Populations in Zebrafish Reveals Gene Regulatory Logic Encoded in the Nuclear Transcriptome
title_full_unstemmed Biotagging of Specific Cell Populations in Zebrafish Reveals Gene Regulatory Logic Encoded in the Nuclear Transcriptome
title_short Biotagging of Specific Cell Populations in Zebrafish Reveals Gene Regulatory Logic Encoded in the Nuclear Transcriptome
title_sort biotagging of specific cell populations in zebrafish reveals gene regulatory logic encoded in the nuclear transcriptome
url http://www.sciencedirect.com/science/article/pii/S2211124717303911
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