Bioengineering bacterial encapsulin nanocompartments as targeted drug delivery system
| Autor: | Kenth Gustafsson, Rana Khalife, Konstantin Pildish, Rupali Dabas, Sofia Esteban Serna, Vladimir Kalinovskiy, Noelle Colant, Matas Deveikis, Alexander Van de Steen, Saverio Charalambous, Hasan Mustafa Khan, Stefanie Frank, Clare M. Robinson, Diana A. Catana |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
QH301-705.5
His6 Hexahistidine Encapsulin Drug delivery system Biomedical Engineering NPs NanoParticles rTurboGFP recombinant Turbo Green Fluorescent Protein Applied Microbiology and Biotechnology Article law.invention iLOV improved Light Oxygen or Voltage-sensing flavoprotein mScarlet a bright monomeric red fluorescent protein STII StrepII-tag an eight-residue peptide sequence (Trp-Ser-His-Pro-Gln-Phe-Glu-Lys) with intrinsic affinity toward streptavidin that can be fused to recombinant protein in various fashions Nanocages DDS Drug Delivery System Structural Biology In vivo law SK-BR-3 Sloan-Kettering Breast cancer cell line/HER2-overexpressing human breast cancer cell line Genetics Cytotoxic protein Annexin V-FITC Annexin V-Fluorescein IsoThiocyanate Conjugate T. maritima Thermotoga maritima Biology (General) HER2 Human Epidermal growth factor Receptor 2 DARPin9.29 Designed Ankyrin Repeat Protein 9.29 biology Chemistry iGEM international Genetically Engineered Machine EPR Enhanced Permeability and Retention effect biology.organism_classification Fusion protein Cell biology DARPin Targeted drug delivery Thermotoga maritima Drug delivery miniSOG mini-Singlet Oxygen Generator Recombinant DNA MSCs Mesenchymal Stem Cells VLPs Virus-Like Particle TP248.13-248.65 Biotechnology |
| Zdroj: | Synthetic and Systems Biotechnology, Vol 6, Iss 3, Pp 231-241 (2021) Synthetic and Systems Biotechnology |
| Popis: | The development of Drug Delivery Systems (DDS) has led to increasingly efficient therapies for the treatment and detection of various diseases. DDS use a range of nanoscale delivery platforms produced from polymeric of inorganic materials, such as micelles, and metal and polymeric nanoparticles, but their variant chemical composition make alterations to their size, shape, or structures inherently complex. Genetically encoded protein nanocages are highly promising DDS candidates because of their modular composition, ease of recombinant production in a range of hosts, control over assembly and loading of cargo molecules and biodegradability. One example of naturally occurring nanocompartments are encapsulins, recently discovered bacterial organelles that have been shown to be reprogrammable as nanobioreactors and vaccine candidates. Here we report the design and application of a targeted DDS platform based on the Thermotoga maritima encapsulin reprogrammed to display an antibody mimic protein called Designed Ankyrin repeat protein (DARPin) on the outer surface and to encapsulate a cytotoxic payload. The DARPin9.29 chosen in this study specifically binds to human epidermal growth factor receptor 2 (HER2) on breast cancer cells, as demonstrated in an in vitro cell culture model. The encapsulin-based DDS is assembled in one step in vivo by co-expressing the encapsulin-DARPin9.29 fusion protein with an engineered flavin-binding protein mini-singlet oxygen generator (MiniSOG), from a single plasmid in Escherichia coli. Purified encapsulin-DARPin_miniSOG nanocompartments bind specifically to HER2 positive breast cancer cells and trigger apoptosis, indicating that the system is functional and specific. The DDS is modular and has the potential to form the basis of a multi-receptor targeted system by utilising the DARPin screening libraries, allowing use of new DARPins of known specificities, and through the proven flexibility of the encapsulin cargo loading mechanism, allowing selection of cargo proteins of choice. |
| Databáze: | OpenAIRE |
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