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Updated: Jan 26, 2026

High Throughput Microinjections of Sea Urchin Zygotes
Published on: January 21, 2014
Generation, expression and utilization of single-domain antibodies for in vivo protein localization and manipulation
Catherine S Schrankel1, Tufan Gökirmak1, Chang-Wook Lee2
1Marine Biology Research Division, Scripps Institution of Oceanography, University of California San Diego, San Diego, CA, United States.
Abstract:
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.
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