1 research outputs found
Exploiting the Protein Corona from Cell Lysate on DNA Functionalized Gold Nanoparticles for Enhanced mRNA Translation
This
study describes the use of DNA functionalized gold nanoparticles (AuNPs)
to enhance the synthesis of proteins in cell lysate and examines the
mechanisms behind the enhanced mRNA translation. With an appropriate
DNA oligomer sequence that hybridizes to the 3′-untranslated
region of two mRNA of interest, insulin and green fluorescent protein
(GFP), we found that these DNA conjugated AuNPs (AuNP-DNA) introduced
into HeLa cell lysate enhanced the synthesis of insulin and GFP by
up to 2.18 and 1.80-fold, respectively, over baseline production with
just the mRNA present. The insulin synthesis was markedly reduced
with non-DNA citrate-capped AuNP (1.25-fold) and AuNP-DNA with a nonspecific
polyÂ(T) sequence (1.25-fold). We showed that both nonspecific adsorption
of ribosomes and translation factors to form a lysate protein corona
on AuNP-DNA and weak hybridization between DNA oligomers and mRNA
of interest were important factors that brought translation factors,
ribosomes, and mRNA into close proximity of each other. This could
reduce the recycling time of ribosomes during mRNA translation, thereby
increasing the efficiency of protein synthesis. The outcome of this
work shows that with rational DNA design, it could be possible to
modulate intracellular biological processes with AuNP-DNA and increase
their production of proteins for various biomedical applications