The Research Progress of Nanobody in The Application of Red Light-Switchable Protein Dimerization Systems

Protein dimerization systems that can be controlled by red light with increased tissue penetration depth are a highly needed tool for clinical applications such as cell and gene therapies. However, existing red light-induced dimerization systems are all based on phytochrome photoreceptors and naturally occurring binding partners with complex structures and nonoptimal in vivo performance limiting mammalian applications. Here, scientist from TIO, and his partner from University of Washington introduce an efficient, generalizable method (COMBINES-LID) for creating highly specific light-induced dimerization systems. It involves a two-step binder screening (phage display and yeast two-hybrid) of a combinatorial nanobody library to obtain binders only binding to the light form of a photo-switchable protein or domain not the dark form. A proof-of-principle was provided by creating nano-body-based, red light-induced dimerization (nanoReD) systems comprising a truncated bacterial phytochrome sensory module using a mammalian endogenous chromophore, biliverdin, and a light-form specific nanobody. Selected nanoReD systems were biochemical characterized and found with low dark activity and high light induction specificity for in vivo activation of gene expression. This work opens new opportunities for creating genetically encoded actuators for the optical manipulation of biological processes.

The research results were published in "ACS Synthetic Biology" in December 2020. The title is "Creating Red Light-Controlled Protein Dimerization Systems as Genetically Encoded Actuators with High Specificity”. The co-first authors of this work is Li Zengpeng from the Third Institute of Oceanography, Ministry of Natural Resources and Dr. Gu Liangcai, from University of Washington is the corresponding author.

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