NaNuTrap: a technique for in vivo cell nucleus labelling using nanobodies
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.
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.Attached Files
Published - dev199822.pdf
Supplemental Material - dev199822supp.pdf
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Additional details
- Eprint ID
- 110320
- Resolver ID
- CaltechAUTHORS:20210820-003328168
- NIH
- R35GM118146
- American Heart Association
- 18POST34080493
- Caltech
- Created
-
2021-08-20Created from EPrint's datestamp field
- Updated
-
2021-09-13Created from EPrint's last_modified field
- Caltech groups
- Division of Biology and Biological Engineering