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Cell Surface Labeling by Engineered Extracellular Vesicles by Hamilton, Nicklas; Claudio, Natalie M.; Armstrong, Randall J.; Pucci, Ferdinando is a Engineering article available to read on EtoBox.

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## Abstract Extracellular vesicles (EVs) can mediate local and long‐range intercellular communication via cell surface signaling. In order to perform in vivo studies of unmanipulated, endogenously released EVs, sensitive but stringent approaches able to detect EV–cell surface interactions are needed. However, isolation and reinfusion of EVs can introduce biases. A rigorous way to study EVs in vivo is by genetically engineering membrane‐bound reporters into parental cells. Still, the amount of reporter molecules that EVs can carry is relatively small, and thus, the sensitivity of the approach is suboptimal. This work addresses this issue by engineering EVs to display a membrane‐bound form of Sortase A (SrtA), a bacterial transpeptidase that can catalyze the transfer of reporter molecules on the much bigger surface of EV‐binding cells. SrtA design and reaction requirements are optimized and validated. Efficient in vitro labeling of EV‐binding cells is achieved, even in the presence of only one N‐terminal glycine on cell surface proteins. As compared to indirect labeling of EV‐binding cells (e.g., using CD63‐GFP fusion), the SrtA‐based approach shows 1–2 log increase in sensitivity, d

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Author

Hamilton, Nicklas; Claudio, Natalie M.; Armstrong, Randall J.; Pucci, Ferdinando

Publisher

Wiley (John Wiley & Sons); Wiley-VCH Verlag; Wiley (ISSN 2366-7478)

Published

2020

Language

EN

Field

Engineering (Physical Sciences)