A genetically encoded probe for imaging nascent and mature HA-tagged proteins in vivo.

Abstract:

:To expand the toolbox of imaging in living cells, we have engineered a single-chain variable fragment binding the linear HA epitope with high affinity and specificity in vivo. The resulting probe, called the HA frankenbody, can light up in multiple colors HA-tagged nuclear, cytoplasmic, membrane, and mitochondrial proteins in diverse cell types. The HA frankenbody also enables state-of-the-art single-molecule experiments in living cells, which we demonstrate by tracking single HA-tagged histones in U2OS cells and single mRNA translation dynamics in both U2OS cells and neurons. Together with the SunTag, we also track two mRNA species simultaneously to demonstrate comparative single-molecule studies of translation can now be done with genetically encoded tools alone. Finally, we use the HA frankenbody to precisely quantify the expression of HA-tagged proteins in developing zebrafish embryos. The versatility of the HA frankenbody makes it a powerful tool for imaging protein dynamics in vivo.

journal_name

Nat Commun

journal_title

Nature communications

authors

Zhao N,Kamijo K,Fox PD,Oda H,Morisaki T,Sato Y,Kimura H,Stasevich TJ

doi

10.1038/s41467-019-10846-1

subject

Has Abstract

pub_date

2019-07-03 00:00:00

pages

2947

issue

1

issn

2041-1723

pii

10.1038/s41467-019-10846-1

journal_volume

10

pub_type

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