ac7b02102_si_001.pdf (1.05 MB)
Antibody-Templated Assembly of an RNA Mimic of Green Fluorescent Protein
journal contribution
posted on 2017-11-13, 00:00 authored by Alessandro Bertucci, Alessandro Porchetta, Francesco RicciOne
of the most intriguing ways through which nature achieves regulation
of biological pathways encompasses the coordination of noncovalent
interactions that bring biomolecules to be colocalized in a designated
restricted space. Inspired by this mechanism, we have explored the
possibility of using antibodies as bivalent biomolecular substrates
for the templated assembly of a functional RNA structure. We have
developed a biosupramolecular complementation assay by assembling
a fluorescent Spinach aptamer, which is a synthetic RNA mimic of the
Green Fluorescent Protein, from its split segments. We have employed
two antigen-tagged RNA strands that, upon binding to the target antibody,
are colocalized in a confined space and can reassemble into the native
Spinach conformation, yielding a measurable fluorescence emission
as a function of the templating antibody concentration. We have demonstrated
the generality of our approach using two different antigen/antibody
systems and found that both platforms show high binding affinity,
specificity for the target antibody, and enough selectivity to work
in crude cellular extracts. This study highlights the potential of
biosupramolecular RNA engineering for the development of innovative
biomimetic tools for nanobiotechnology and bioanalytical assays.