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Methods Cell Biol
2019 Jan 01;151:353-376. doi: 10.1016/bs.mcb.2018.11.005.
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Generation, expression and utilization of single-domain antibodies for in vivo protein localization and manipulation in sea urchin embryos.
Schrankel CS
,
Gökirmak T
,
Lee CW
,
Chang G
,
Hamdoun A
.
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Single-domain antibodies, also known as nanobodies, are small antigen-binding fragments (~15kDa) that are derived from heavy chain only antibodies present in camelids (VHH, from camels and llamas), and cartilaginous fishes (VNAR, from sharks). Nanobody V-like domains are useful alternatives to conventional antibodies due to their small size, and high solubility and stability across many applications. In addition, phage display, ribosome display, and mRNA/cDNA display methods can be used for the efficient generation and optimization of binders in vitro. The resulting nanobodies can be genetically encoded, tagged, and expressed in cells for in vivo localization and functional studies of target proteins. Collectively, these properties make nanobodies ideal for use within echinoderm embryos. This chapter describes the optimization and imaging of genetically encoded nanobodies in the sea urchin embryo. Examples of live-cell antigen tagging (LCAT) and the manipulation of green fluorescent protein (GFP) are shown. We discuss the potentially transformative applications of nanobody technology for probing membrane protein trafficking, cytoskeleton re-organization, receptor signaling events, and gene regulation during echinoderm development.
Adams,
Synthetic antibody technologies.
2014,
Pubmed
Anton,
Site-specific recruitment of epigenetic factors with a modular CRISPR/Cas system.
2017,
Pubmed
Barsi,
Genome-wide assessment of differential effector gene use in embryogenesis.
2015,
Pubmed
,
Echinobase
Beghein,
Nanobody Technology: A Versatile Toolkit for Microscopic Imaging, Protein-Protein Interaction Analysis, and Protein Function Exploration.
2017,
Pubmed
Bothma,
LlamaTags: A Versatile Tool to Image Transcription Factor Dynamics in Live Embryos.
2018,
Pubmed
Böldicke,
Single domain antibodies for the knockdown of cytosolic and nuclear proteins.
2017,
Pubmed
Campanale,
Migration of sea urchin primordial germ cells.
2014,
Pubmed
,
Echinobase
Caussinus,
Fluorescent fusion protein knockout mediated by anti-GFP nanobody.
2011,
Pubmed
Dmitriev,
Nanobodies as Probes for Protein Dynamics in Vitro and in Cells.
2016,
Pubmed
Doshi,
In vitro nanobody discovery for integral membrane protein targets.
2014,
Pubmed
Fridy,
A robust pipeline for rapid production of versatile nanobody repertoires.
2014,
Pubmed
Greenberg,
A new antigen receptor gene family that undergoes rearrangement and extensive somatic diversification in sharks.
1995,
Pubmed
Gökirmak,
Localization and substrate selectivity of sea urchin multidrug (MDR) efflux transporters.
2012,
Pubmed
,
Echinobase
Hamers-Casterman,
Naturally occurring antibodies devoid of light chains.
1993,
Pubmed
Harmansa,
Dpp spreading is required for medial but not for lateral wing disc growth.
2015,
Pubmed
Harmansa,
A nanobody-based toolset to investigate the role of protein localization and dispersal in Drosophila.
2017,
Pubmed
Harmansa,
Protein binders and their applications in developmental biology.
2018,
Pubmed
Helma,
Nanobodies and recombinant binders in cell biology.
2015,
Pubmed
Kirchhofer,
Modulation of protein properties in living cells using nanobodies.
2010,
Pubmed
Lepage,
Expression of exogenous mRNAs to study gene function in the sea urchin embryo.
2004,
Pubmed
,
Echinobase
Maier,
Real-time analysis of epithelial-mesenchymal transition using fluorescent single-domain antibodies.
2015,
Pubmed
McMahon,
Yeast surface display platform for rapid discovery of conformationally selective nanobodies.
2018,
Pubmed
Moutel,
NaLi-H1: A universal synthetic library of humanized nanobodies providing highly functional antibodies and intrabodies.
2016,
Pubmed
Muyldermans,
Nanobodies: natural single-domain antibodies.
2013,
Pubmed
Newnham,
Functional inhibition of β-catenin-mediated Wnt signaling by intracellular VHH antibodies.
2015,
Pubmed
Panza,
Live imaging of endogenous protein dynamics in zebrafish using chromobodies.
2015,
Pubmed
Rasmussen,
Structure of a nanobody-stabilized active state of the β(2) adrenoceptor.
2011,
Pubmed
Rast,
Transgenic manipulation of the sea urchin embryo.
2000,
Pubmed
,
Echinobase
Riedl,
Lifeact: a versatile marker to visualize F-actin.
2008,
Pubmed
Roberts,
RNA-peptide fusions for the in vitro selection of peptides and proteins.
1997,
Pubmed
Rothbauer,
Targeting and tracing antigens in live cells with fluorescent nanobodies.
2006,
Pubmed
Schindelin,
Fiji: an open-source platform for biological-image analysis.
2012,
Pubmed
Shin,
Nanobody-targeted E3-ubiquitin ligase complex degrades nuclear proteins.
2015,
Pubmed
Shipp,
A teaching laboratory on the activation of xenobiotic transporters at fertilization of sea urchins.
2019,
Pubmed
,
Echinobase
Stanfield,
Crystal structure of a shark single-domain antibody V region in complex with lysozyme.
2004,
Pubmed
Tang,
A nanobody-based system using fluorescent proteins as scaffolds for cell-specific gene manipulation.
2013,
Pubmed
Van Impe,
A nanobody targeting the F-actin capping protein CapG restrains breast cancer metastasis.
2013,
Pubmed
Wada,
Development of Next-Generation Peptide Binders Using In vitro Display Technologies and Their Potential Applications.
2013,
Pubmed
Yamagata,
Reporter-nanobody fusions (RANbodies) as versatile, small, sensitive immunohistochemical reagents.
2018,
Pubmed
Zuo,
Institute collection and analysis of Nanobodies (iCAN): a comprehensive database and analysis platform for nanobodies.
2017,
Pubmed
