Version 1
: Received: 10 October 2023 / Approved: 11 October 2023 / Online: 12 October 2023 (05:31:34 CEST)
How to cite:
JIA, Y.; Bleicher, F.; Merabet, S. Current Insights and Perspectives on High-Throughput BiFC. Preprints2023, 2023100764. https://doi.org/10.20944/preprints202310.0764.v1
JIA, Y.; Bleicher, F.; Merabet, S. Current Insights and Perspectives on High-Throughput BiFC. Preprints 2023, 2023100764. https://doi.org/10.20944/preprints202310.0764.v1
JIA, Y.; Bleicher, F.; Merabet, S. Current Insights and Perspectives on High-Throughput BiFC. Preprints2023, 2023100764. https://doi.org/10.20944/preprints202310.0764.v1
APA Style
JIA, Y., Bleicher, F., & Merabet, S. (2023). Current Insights and Perspectives on High-Throughput BiFC. Preprints. https://doi.org/10.20944/preprints202310.0764.v1
Chicago/Turabian Style
JIA, Y., Françoise Bleicher and Samir Merabet. 2023 "Current Insights and Perspectives on High-Throughput BiFC" Preprints. https://doi.org/10.20944/preprints202310.0764.v1
Abstract
For a comprehensive exploration of the Bimolecular Fluorescence Complementation (BiFC) approach, we critically assess whether protein pairs establish direct or indirect interactions within this context. Emphasizing the multifaceted nature of protein interactions, this review underscores the imperative of delineating macromolecular complex characteristics across five distinct levels: composition, stoichiometry, copy number, topology, and dynamics. A pivotal discussion revolves around the challenges introduced by the overexpression of target proteins and the potential artificial ramifications of fusion proteins. The significance of stoichiometry in protein interactions is illuminated, with the HOXA9/PBX1 interaction serving as a pertinent case study. The research identifies and elaborates on the inherent limitations of prevailing large-scale BiFC screenings, advocating for the incorporation of rigorous negative controls. A novel proposition is introduced in the form of a high-throughput multicolor BiFC strategy, exemplified using the HOXA9 and PBX1 proteins. The paper culminates with an insightful discourse on the implications of tags within the BiFC system, presenting the TriFC (Tripartite Fluorescence Complementation) assay as a promising alternative.
Keywords
HT-BiFC; HOXA9/PBX1; PPI
Subject
Biology and Life Sciences, Biology and Biotechnology
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
