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Green fluorescent protein (GFP): is seeing believing and is that enough?

journal contribution
posted on 2023-07-26, 14:17 authored by Susan A. Shorter, Marie W. Pettit, Paul D. R. Dyer, Emma Coakley Youngs, Monique A. M. Gorringe-Pattrick, Samer El-Daher, Simon Richardson
Intracellular compartmentalisation is a significant barrier to the successful nucleocytosolic delivery of biologics. The endocytic system has been shown to be responsible for compartmentalisation, providing an entry point, and trigger(s) for the activation of drug delivery systems. Consequently, many of the technologies used to understand endocytosis have found utility within the field of drug delivery. The use of fluorescent proteins as markers denoting compartmentalisation within the endocytic system has become commonplace. Several of the limitations associated with the use of green fluorescent protein (GFP) within the context of drug delivery have been explored here by asking a series of related questions: (1) Are molecules that regulate fusion to a specific compartment (i.e. Rab- or SNARE-GFP fusions) a good choice of marker for that compartment? (2) How reliable was GFP-marker overexpression when used to define a given endocytic compartment? (3) Can glutathione-s-transferase (GST) fused in frame with GFP (GST–GFP) act as a fluid phase endocytic probe? (4) Was GFP fluorescence a robust indicator of (GFP) protein integrity? This study concluded that there are many appropriate and useful applications for GFP; however, thought and an understanding of the biological and physicochemical character of these markers are required for the generation of meaningful data.

History

Refereed

  • Yes

Volume

25

Issue number

9-10

Page range

809-817

Publication title

Journal of Drug Targeting

ISSN

1029-2330

Publisher

Taylor & Francis

Language

  • other

Legacy posted date

2018-03-19

Legacy Faculty/School/Department

ARCHIVED Faculty of Science & Technology (until September 2018)

Note

An author accepted version of this paper available through University of Greenwich's repository : http://gala.gre.ac.uk/16592/

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