PT  - JOURNAL ARTICLE
AU  - Diogo Bessa-Neto
AU  - Alexander Kuhlemann
AU  - Gerti Beliu
AU  - Valeria Pecoraro
AU  - Sören Doose
AU  - Natacha Retailleau
AU  - Nicolas Chevrier
AU  - David Perrais
AU  - Markus Sauer
AU  - Daniel Choquet
TI  - Bioorthogonal labeling of transmembrane proteins with non-canonical amino acids allows access to masked epitopes in live neurons
AID  - 10.1101/2021.02.27.433189
DP  - 2021 Jan 01
TA  - bioRxiv
PG  - 2021.02.27.433189
4099  - http://biorxiv.org/content/early/2021/02/28/2021.02.27.433189.short
4100  - http://biorxiv.org/content/early/2021/02/28/2021.02.27.433189.full
AB  - Progress in biological imaging is intrinsically linked to advances in labeling methods. The explosion in the development of high-resolution and super-resolution imaging calls for new approaches to label targets with small probes. These should allow to faithfully report the localization of the target within the imaging resolution – typically nowadays a few nanometers - and allow access to any epitope of the target, in the native cellular and tissue environment. We report here the development of a complete labeling and imaging pipeline using genetic code expansion and non-canonical amino acids in primary neurons that allows to fluorescently label masked epitopes in target transmembrane proteins in live neurons, both in dissociated culture and organotypic brain slices. This allowed us to image the differential localization of two glutamate receptor auxiliary proteins in complex with their partner with a variety of methods including widefield, confocal, and dSTORM super-resolution microscopy.Competing Interest StatementThe authors have declared no competing interest.