CaltechAUTHORS
  A Caltech Library Service

NaNuTrap: a technique for in vivo cell nucleus labelling using nanobodies

Ákos, Zsuzsa and Dunipace, Leslie and Stathopoulos, Angelike (2021) NaNuTrap: a technique for in vivo cell nucleus labelling using nanobodies. Development, 148 (18). Art. No. dev199822. ISSN 0950-1991. doi:10.1242/dev.199822. https://resolver.caltech.edu/CaltechAUTHORS:20210820-003328168

[img] PDF - Published Version
Creative Commons Attribution.

5MB
[img] PDF - Supplemental Material
Creative Commons Attribution.

3MB

Use this Persistent URL to link to this item: https://resolver.caltech.edu/CaltechAUTHORS:20210820-003328168

Abstract

In vivo cell labelling is challenging in fast developmental processes because many cell types differentiate more quickly than the maturation time of fluorescent proteins, making visualization of these tissues impossible with standard techniques. Here, we present a nanobody-based method, Nanobody Nuclear Trap (NaNuTrap), which works with the existing Gal4/UAS system in Drosophila and allows for early in vivo cell nuclei labelling independently of the maturation time of the fluorescent protein. This restores the utility of fluorescent proteins that have longer maturation times, such as those used in two-photon imaging, for live imaging of fast or very early developmental processes. We also present a more general application of this system, whereby NaNuTrap can convert cytoplasmic GFP expressed in any existing transgenic fly line into a nuclear label. This nuclear re-localization of the fluorescent signal can improve the utility of the GFP label, e.g. in cell counting, as well as resulting in a general increase in intensity of the live fluorescent signal. We demonstrate these capabilities of NaNuTrap by effectively tracking subsets of cells during the fast movements associated with gastrulation.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1242/dev.199822DOIArticle
ORCID:
AuthorORCID
Ákos, Zsuzsa0000-0002-4587-6212
Dunipace, Leslie0000-0002-1739-6537
Stathopoulos, Angelike0000-0001-6597-2036
Additional Information:© 2021. Published by The Company of Biologists Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed. Received 24 May 2021; Accepted 12 July 2021. We thank Jacques Bothma for helpful discussion, the Hernan Garcia lab for fly lines and technical advice, the Caltech Beckman Imaging centre for support, Frank Macabenta for helping with schematics, Susan Newcomb for comments on the manuscript, and the anonymous reviewers for helpful suggestions. This work was funded by the National Institutes of Health (R35GM118146 to A.S.) and by the American Heart Association (18POST34080493 to Z.Á.). Open Access funding provided by California Institute Of Technology. Deposited in PMC for immediate release. Author contributions: Conceptualization: Z.Á., L.D., A.S.; Methodology: Z.Á., L.D.; Validation: Z.Á., A.S.; Formal analysis: Z.Á.; Investigation: Z.Á.; Resources: A.S.; Data curation: Z.Á.; Writing - original draft: Z.Á.; Writing - review & editing: Z.Á., L.D., A.S.; Visualization: Z.Á.; Supervision: A.S.; Project administration: A.S.; Funding acquisition: Z.Á., A.S. The authors declare no competing or financial interests.
Funders:
Funding AgencyGrant Number
NIHR35GM118146
American Heart Association18POST34080493
CaltechUNSPECIFIED
Subject Keywords:Nanobody, Nucleus, GFP maturation, Live in vivo imaging, Cell tracking, Drosophila melanogaster
Issue or Number:18
DOI:10.1242/dev.199822
Record Number:CaltechAUTHORS:20210820-003328168
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20210820-003328168
Official Citation:Zsuzsa Ákos, Leslie Dunipace, Angelike Stathopoulos; NaNuTrap: a technique for in vivo cell nucleus labelling using nanobodies. Development 15 September 2021; 148 (18): dev199822. doi: https://doi.org/10.1242/dev.199822
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:110320
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:20 Aug 2021 16:00
Last Modified:13 Sep 2021 20:21

Repository Staff Only: item control page