title: Exploiting the Protein Corona from Cell Lysate on DNA Functionalized Gold Nanoparticles for Enhanced mRNA Translation.
abstract: 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 poly(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.
authors.affiliation: Department of Biomedical Engineering, National University of Singapore , Singapore 117583.|Bioprocessing Technology Institute, Agency for Science, Technology and Research , Singapore 138668.|NUS Graduate School for Integrative Sciences and Engineering, National University of Singapore , Singapore 117456.
authors.forename: Kian Ping
authors.initials: KP
authors.lastname: Chan
authors.affiliation: Department of Biomedical Engineering, National University of Singapore , Singapore 117583.
authors.forename: Yang
authors.initials: Y
authors.lastname: Gao
authors.affiliation: Department of Biomedical Engineering, National University of Singapore , Singapore 117583.
authors.forename: Jeremy Xianwei
authors.initials: JX
authors.lastname: Goh
authors.affiliation: Faculty of Science, National University of Singapore , Singapore 117546.
authors.forename: Dewi
authors.initials: D
authors.lastname: Susanti
authors.affiliation: Department of Biomedical Engineering, National University of Singapore , Singapore 117583.
authors.forename: Eugenia Li Ling
authors.initials: EL
authors.lastname: Yeo
authors.affiliation: Bioprocessing Technology Institute, Agency for Science, Technology and Research , Singapore 138668.|Department of Microbiology and Immunology, National University of Singapore , Singapore 117545.
authors.forename: Sheng-Hao
authors.initials: SH
authors.lastname: Chao
authors.affiliation: Department of Biomedical Engineering, National University of Singapore , Singapore 117583.|NUS Graduate School for Integrative Sciences and Engineering, National University of Singapore , Singapore 117456.
authors.forename: James Chen Yong
authors.identifier: http://orcid.org/0000-0002-2247-6929
authors.initials: JC
authors.lastname: Kah
mesh_terms: D004247:DNA; D006046:Gold; D006367:HeLa Cells; D006801:Humans; D053768:Metal Nanoparticles; D014176:Protein Biosynthesis; D000066970:Protein Corona / Q000737:chemistry*
chemical_list: D000066970:Protein Corona; D006046:Gold; D004247:DNA
keywords: DNA; gold nanoparticles; hybridization; mRNA translation; nonspecific adsorption; protein corona