| Autor: |
Trashi I; Department of Chemistry and Biochemistry, The University of Texas at Dallas, Richardson, Texas 75080, USA. gassensmith@utdallas.edu., Durbacz MZ; Department of Molecular Biology and Hamon Center for Regenerative Science and Medicine, University of Texas Southwestern Medical Center, Dallas, Texas, USA., Trashi O; Department of Chemistry and Biochemistry, The University of Texas at Dallas, Richardson, Texas 75080, USA. gassensmith@utdallas.edu., Wijesundara YH; Department of Chemistry and Biochemistry, The University of Texas at Dallas, Richardson, Texas 75080, USA. gassensmith@utdallas.edu., Ehrman RN; Department of Chemistry and Biochemistry, The University of Texas at Dallas, Richardson, Texas 75080, USA. gassensmith@utdallas.edu., Chiev AC; Department of Chemistry and Biochemistry, The University of Texas at Dallas, Richardson, Texas 75080, USA. gassensmith@utdallas.edu., Darwin CB; Department of Chemistry and Biochemistry, The University of Texas at Dallas, Richardson, Texas 75080, USA. gassensmith@utdallas.edu., Herbert FC; Department of Chemistry and Biochemistry, The University of Texas at Dallas, Richardson, Texas 75080, USA. gassensmith@utdallas.edu., Gadhvi J; Department of Biological Science, The University of Texas at Dallas, Richardson, Texas 75080, USA., De Nisco NJ; Department of Biological Science, The University of Texas at Dallas, Richardson, Texas 75080, USA., Nielsen SO; Department of Chemistry and Biochemistry, The University of Texas at Dallas, Richardson, Texas 75080, USA. gassensmith@utdallas.edu., Gassensmith JJ; Department of Chemistry and Biochemistry, The University of Texas at Dallas, Richardson, Texas 75080, USA. gassensmith@utdallas.edu.; Department of Bioengineering, The University of Texas at Dallas, Richardson, Texas 75080, USA. |
| Abstrakt: |
Virus-like particles (VLPs) are engineered nanoparticles that mimic the properties of viruses-like high tolerance to heat and proteases-but lack a viral genome, making them non-infectious. They are easily modified chemically and genetically, making them useful in drug delivery, enhancing vaccine efficacy, gene delivery, and cancer immunotherapy. One such VLP is Qβ, which has an affinity towards an RNA hairpin structure found in its viral RNA that drives the self-assembly of the capsid. It is possible to usurp the native way infectious Qβ self-assembles to encapsidate its RNA to place enzymes inside the VLP's lumen as a protease-resistant cage. Further, using RNA templates that mimic the natural self-assembly of the native capsid, fluorescent proteins (FPs) have been placed inside VLPs in a "one pot" expression system. Autofluorescence in tissues can lead to misinterpretation of results and unreliable science, so we created a single-pot expression system that uses the fluorescent protein smURFP, which avoids autofluorescence and has spectral properties compatible with standard commercial filter sets on confocal microscopes. In this work, we were able to simplify the existing "one-pot" expression system while creating high-yielding fluorescent VLP nanoparticles that could easily be imaged inside lung epithelial tissue. |
