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The response of the adaptive immune system is augmented by multimeric presentation of a specific antigen, resembling viral particles. Several vaccines have been designed based on natural or designed protein scaffolds, which exhibited a potent adaptive immune response to antigens; however, antibodies are also generated against the scaffold, which may impair subsequent vaccination. In order to compare polypeptide scaffolds of different size and oligomerization state with respect to their efficiency, including anti-scaffold immunity, we compared several strategies of presentation of the RBD domain of the SARS-CoV-2 spike protein, an antigen aiming to generate neutralizing antibodies. A comparison of several genetic fusions of RBD to different nanoscaffolding domains (foldon, ferritin, lumazine synthase, and β-annulus peptide) delivered as DNA plasmids demonstrated a strongly augmented immune response, with high titers of neutralizing antibodies and a robust T-cell response in mice. Antibody titers and virus neutralization were most potently enhanced by fusion to the small β-annulus peptide scaffold, which itself triggered a minimal response in contrast to larger scaffolds. The β-annulus fused RBD protein increased residence in lymph nodes and triggered the most potent viral neutralization in immunization by a recombinant protein. Results of the study support the use of a nanoscaffolding platform using the β-annulus peptide for vaccine design.
Duško Lainšček; Tina Fink; Vida Forstnerič; Iva Hafner-Bratkovič; Sara Orehek; Žiga Strmšek; Mateja Manček-Keber; Peter Pečan; Hana Esih; Špela Malenšek; Jana Aupič; Petra Dekleva; Tjaša Plaper; Sara Vidmar; Lucija Kadunc; Mojca Benčina; Neža Omersa; Gregor Anderluh; Florence Pojer; Kelvin Lau; David Hacker; Bruno Correia; David Peterhoff; Ralf Wagner; Valter Bergant; Alexander Herrmann; Andreas Pichlmair; Roman Jerala. A Nanoscaffolded Spike-RBD Vaccine Provides Protection against SARS-CoV-2 with Minimal Anti-Scaffold Response. Vaccines 2021, 9, 431 .
AMA StyleDuško Lainšček, Tina Fink, Vida Forstnerič, Iva Hafner-Bratkovič, Sara Orehek, Žiga Strmšek, Mateja Manček-Keber, Peter Pečan, Hana Esih, Špela Malenšek, Jana Aupič, Petra Dekleva, Tjaša Plaper, Sara Vidmar, Lucija Kadunc, Mojca Benčina, Neža Omersa, Gregor Anderluh, Florence Pojer, Kelvin Lau, David Hacker, Bruno Correia, David Peterhoff, Ralf Wagner, Valter Bergant, Alexander Herrmann, Andreas Pichlmair, Roman Jerala. A Nanoscaffolded Spike-RBD Vaccine Provides Protection against SARS-CoV-2 with Minimal Anti-Scaffold Response. Vaccines. 2021; 9 (5):431.
Chicago/Turabian StyleDuško Lainšček; Tina Fink; Vida Forstnerič; Iva Hafner-Bratkovič; Sara Orehek; Žiga Strmšek; Mateja Manček-Keber; Peter Pečan; Hana Esih; Špela Malenšek; Jana Aupič; Petra Dekleva; Tjaša Plaper; Sara Vidmar; Lucija Kadunc; Mojca Benčina; Neža Omersa; Gregor Anderluh; Florence Pojer; Kelvin Lau; David Hacker; Bruno Correia; David Peterhoff; Ralf Wagner; Valter Bergant; Alexander Herrmann; Andreas Pichlmair; Roman Jerala. 2021. "A Nanoscaffolded Spike-RBD Vaccine Provides Protection against SARS-CoV-2 with Minimal Anti-Scaffold Response." Vaccines 9, no. 5: 431.
The contribution of ribosome heterogeneity and ribosome-associated proteins to the molecular control of proteomes in health and disease remains unclear. Here, we demonstrate that survival motor neuron (SMN) protein—the loss of which causes the neuromuscular disease spinal muscular atrophy (SMA)—binds to ribosomes and that this interaction is tissue-dependent. SMN-primed ribosomes are preferentially positioned within the first five codons of a set of mRNAs that are enriched for translational enhancer sequences in the 5′ untranslated region (UTR) and rare codons at the beginning of their coding sequence. These SMN-specific mRNAs are associated with neurogenesis, lipid metabolism, ubiquitination, chromatin regulation and translation. Loss of SMN induces ribosome depletion, especially at the beginning of the coding sequence of SMN-specific mRNAs, leading to impairment of proteins that are involved in motor neuron function and stability, including acetylcholinesterase. Thus, SMN plays a crucial role in the regulation of ribosome fluxes along mRNAs encoding proteins that are relevant to SMA pathogenesis. Lauria et al. show that SMN, the loss of which causes spinal muscular atrophy (SMA), preferentially positions ribosomes within the first five codons of SMA-related mRNAs and enhances their translation.
Fabio Lauria; Paola Bernabò; Toma Tebaldi; Ewout Joan Nicolaas Groen; Elena Perenthaler; Federica Maniscalco; Annalisa Rossi; Deborah Donzel; Massimiliano Clamer; Marta Marchioretto; Neža Omersa; Julia Orri; Mauro Dalla Serra; Gregor Anderluh; Alessandro Quattrone; Alberto Inga; Thomas Henry Gillingwater; Gabriella Viero. SMN-primed ribosomes modulate the translation of transcripts related to spinal muscular atrophy. Nature 2020, 22, 1239 -1251.
AMA StyleFabio Lauria, Paola Bernabò, Toma Tebaldi, Ewout Joan Nicolaas Groen, Elena Perenthaler, Federica Maniscalco, Annalisa Rossi, Deborah Donzel, Massimiliano Clamer, Marta Marchioretto, Neža Omersa, Julia Orri, Mauro Dalla Serra, Gregor Anderluh, Alessandro Quattrone, Alberto Inga, Thomas Henry Gillingwater, Gabriella Viero. SMN-primed ribosomes modulate the translation of transcripts related to spinal muscular atrophy. Nature. 2020; 22 (10):1239-1251.
Chicago/Turabian StyleFabio Lauria; Paola Bernabò; Toma Tebaldi; Ewout Joan Nicolaas Groen; Elena Perenthaler; Federica Maniscalco; Annalisa Rossi; Deborah Donzel; Massimiliano Clamer; Marta Marchioretto; Neža Omersa; Julia Orri; Mauro Dalla Serra; Gregor Anderluh; Alessandro Quattrone; Alberto Inga; Thomas Henry Gillingwater; Gabriella Viero. 2020. "SMN-primed ribosomes modulate the translation of transcripts related to spinal muscular atrophy." Nature 22, no. 10: 1239-1251.
Lipid membranes are becoming increasingly popular in synthetic biology due to their biophysical properties and crucial role in communication between different compartments. Several alluring protein-membrane sensors have already been developed, whereas protein logic gates designs on membrane-embedded proteins are very limited. Here we demonstrate the construction of a two-level protein-membrane logic gate with an OR-AND logic. The system consists of an engineered pH-dependent pore-forming protein listeriolysin O and its DARPin-based inhibitor, conjugated to a lipid vesicle membrane. The gate responds to low pH and removal of the inhibitor from the membrane either by switching to a reducing environment, protease cleavage, or any other signal depending on conjugation chemistry used for inhibitor attachment to the membrane. This unique protein logic gate vesicle system advances generic sensing and actuator platforms used in synthetic biology and could be utilized in drug delivery.
Neža Omersa; Saša Aden; Matic Kisovec; Marjetka Podobnik; Gregor Anderluh. Design of Protein Logic Gate System Operating on Lipid Membranes. ACS Synthetic Biology 2020, 9, 316 -328.
AMA StyleNeža Omersa, Saša Aden, Matic Kisovec, Marjetka Podobnik, Gregor Anderluh. Design of Protein Logic Gate System Operating on Lipid Membranes. ACS Synthetic Biology. 2020; 9 (2):316-328.
Chicago/Turabian StyleNeža Omersa; Saša Aden; Matic Kisovec; Marjetka Podobnik; Gregor Anderluh. 2020. "Design of Protein Logic Gate System Operating on Lipid Membranes." ACS Synthetic Biology 9, no. 2: 316-328.
Perforation of cellular membranes by pore-forming proteins can affect cell physiology, tissue integrity, or immune response. Since many pore-forming proteins are toxins or highly potent virulence factors, they represent an attractive target for the development of molecules that neutralize their actions with high efficacy. There has been an assortment of inhibitors developed to specifically obstruct the activity of pore-forming proteins, in addition to vaccination and antibiotics that serve as a plausible treatment for the majority of diseases caused by bacterial infections. Here we review a wide range of potential inhibitors that can specifically and effectively block the activity of pore-forming proteins, from small molecules to more specific macromolecular systems, such as synthetic nanoparticles, antibodies, antibody mimetics, polyvalent inhibitors, and dominant negative mutants. We discuss their mechanism of inhibition, as well as advantages and disadvantages.
Neža Omersa; Marjetka Podobnik; Gregor Anderluh. Inhibition of Pore-Forming Proteins. Toxins 2019, 11, 545 .
AMA StyleNeža Omersa, Marjetka Podobnik, Gregor Anderluh. Inhibition of Pore-Forming Proteins. Toxins. 2019; 11 (9):545.
Chicago/Turabian StyleNeža Omersa; Marjetka Podobnik; Gregor Anderluh. 2019. "Inhibition of Pore-Forming Proteins." Toxins 11, no. 9: 545.