Engineering a Virus-like Particle to Display Peptide Insertions Using an Apparent Fitness Landscape

Robinson, Stephanie A.; Hartman, Emily C.; Ikwuagwu, Bon; Francis, Matthew B.; Tullman‐Ercek, Danielle

doi:10.1021/acs.biomac.0c00987

Cited by 15 publications

(10 citation statements)

References 41 publications

(88 reference statements)

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“…It will be interesting to extend our model to explicitly incorporate scaffold-mediated cargo-cargo and shellcargo interactions, for example as Mohajerani et al 62 did for a single-component cargo. More broadly, the predictions from our models can be extended beyond microcompartments, for example to re-engineered viral capsids [46][47][48]51,[147][148][149][150][151][152][153][154][155] or synthetic capsids constructed via protein engineering (e.g. [168][169][170] or DNA origami 171 to assemble around specific cargoes.…”

Section: Discussionmentioning

confidence: 99%

“…Although our model is motivated by bacterial microcompartments, it is sufficiently general to describe other proteinaceous shells, including viruses and microcompartments that are reengineered to encapsulate designer cargoes (e.g. 22,[30][31][32][33][34][35][36][37][38][39]41,[46][47][48][50][51][52][147][148][149][150][151][152][153][154][155][156] ).…”

Section: A Model Overviewmentioning

confidence: 99%

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Microcompartment assembly around multicomponent fluid cargoes

Tsidilkovski

Hagan

2022

Preprint

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This article describes dynamical simulations of the assembly of an icosahedral protein shell around a bi- component fluid cargo. Our simulations are motivated by bacterial microcompartments, which are protein shells found in bacteria that assemble around a complex of enzymes and other components involved in certain metabolic processes. The simulations demonstrate that the relative interaction strengths among the different cargo species play a key role in determining the amount of each species that is encapsulated, their spatial organization, and the nature of the shell assembly pathways. However, the shell protein-protein and shell-cargo interactions that help drive assembly and encapsulation also influence cargo composition within certain parameter regimes. These behaviors are governed by combination of thermodynamic and kinetic effects. In addition to elucidating how natural microcompartments encapsulate multiple components involved within reaction cascades, these results have implications for synthetic biology efforts to colocalize alternative sets of molecules within microcompartments to acceleratereactions. More broadly, the results suggest that coupling between self-assembly and multicomponent liquid-liquid phase separation may play a role in organization of the cellular cytoplasm.

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Section: Discussionmentioning

confidence: 99%

Section: A Model Overviewmentioning

confidence: 99%

Microcompartment assembly around multicomponent fluid cargoes

Tsidilkovski

Hagan

2022

Preprint

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“…Bacteriophage MS2 is recombinantly produced using a pBAD plasmid. (Robinson et al, 2020) Transformed single colonies are added to LB media supplemented with appropriate antibiotics (e.g., 50 µg/mL ampicillin) and subsequently induced with arabinose. Bacterial cells are collected via centrifugation and frozen at -80°C.…”

Section: Ms2mentioning

confidence: 99%

Rip It, Stitch It, Click It: A Chemist’s Guide to VLP Manipulation

Herbert

Wijesundara

Kumari

et al. 2022

Preprint

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Viruses are some of nature’s most ubiquitous self-assembled molecular containers. Evolutionary pressures have created some incredibly robust, thermally and enzymatically resistant containers to transport delicate genetic information safely. Virus-like particles (VLPs) are human-engineered non-infectious systems that inherit the parent virus’ ability to self-assemble under controlled conditions while being non-infectious. VLPs and plant-based viral nanoparticles are becoming increasingly popular in medicine as their self-assembly properties are exploitable for applications ranging from diagnostic tools to targeted drug delivery. Understanding the basic structure and principles underlying the assembly of higher-order structures has allowed researchers to disassemble (rip it), functionalize (click it), and reassemble (stitch it) these systems on demand. This review focuses on the current toolbox of strategies developed to manipulate these systems by ripping, stitching, and clicking to create new technologies in the biomedical space.

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“…By inserting all possible three‐residue peptides into the FG loop of MS2 CP, the “systematic mutagenesis and assembled particle selection” method was adopted to elicit the sequencing of the selected peptide insertion library. [ 163 ] Positive controls for molecular assays are used in reverse transcription‐polymerase chain reactions (RT‐PCRs), which serve as a detection tool for COVID‐19 infection to validate each test with high accuracy. Recently, to overcome the related limitations of positive controls, including cold‐chain distribution requirements, a biomimetic virus‐like particle was devised from a bacteriophage and a plant virus to encapsidate a SARS‐CoV‐2 detection module.…”

Section: Nanocarriersmentioning

confidence: 99%

Nanotechnology‐Assisted RNA Delivery: From Nucleic Acid Therapeutics to COVID‐19 Vaccines

et al. 2021

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In recent years, the main quest of science has been the pioneering of the groundbreaking biomedical strategies needed for achieving a personalized medicine. Ribonucleic acids (RNAs) are outstanding bioactive macromolecules identified as pivotal actors in regulating a wide range of biochemical pathways. The ability to intimately control the cell fate and tissue activities makes RNA‐based drugs the most fascinating family of bioactive agents. However, achieving a widespread application of RNA therapeutics in humans is still a challenging feat, due to both the instability of naked RNA and the presence of biological barriers aimed at hindering the entrance of RNA into cells. Recently, material scientists’ enormous efforts have led to the development of various classes of nanostructured carriers customized to overcome these limitations. This work systematically reviews the current advances in developing the next generation of drugs based on nanotechnology‐assisted RNA delivery. The features of the most used RNA molecules are presented, together with the development strategies and properties of nanostructured vehicles. Also provided is an in‐depth overview of various therapeutic applications of the presented systems, including coronavirus disease vaccines and the newest trends in the field. Lastly, emerging challenges and future perspectives for nanotechnology‐mediated RNA therapies are discussed.

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Engineering a Virus-like Particle to Display Peptide Insertions Using an Apparent Fitness Landscape

Cited by 15 publications

References 41 publications

Microcompartment assembly around multicomponent fluid cargoes

Microcompartment assembly around multicomponent fluid cargoes

Rip It, Stitch It, Click It: A Chemist’s Guide to VLP Manipulation

Nanotechnology‐Assisted RNA Delivery: From Nucleic Acid Therapeutics to COVID‐19 Vaccines

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