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Transduction of Human Cells with Polymer-complexed Ecotropic Lentivirus for Enhanced Biosafety
Published on: July 24, 2011
Protein transfer into human cells by VSV-G-induced nanovesicles
Philippe-Emmanuel Mangeot1, Sandra Dollet, Mathilde Girard
1Université de Lyon, INSERM, U851, Lyon, France. philippe.mangeot@inserm.fr
Summary
Researchers developed gesicles, engineered vesicles that deliver proteins into human cells. This novel protein-transduction method enhances safety for stem cell research and offers versatile non-genetic functional delivery.
Area of Science:
- Cell Biology
- Biotechnology
- Molecular Medicine
Background:
- Efficient protein delivery into mammalian cells is crucial for research and therapeutic applications.
- Current methods for exogenous protein expression face limitations in efficiency and safety.
- Developing novel protein-transduction systems is essential for advancing cell-based therapies and biological studies.
Purpose of the Study:
- To investigate the potential of engineered vesicles, termed gesicles, for delivering exogenous proteins into human cells.
- To demonstrate the capacity of gesicles to transport various proteins, including cytoplasmic, nuclear, and surface proteins.
- To evaluate the utility of gesicles for enhancing biosafety in stem cell research.
Main Methods:
- Overexpression of the vesicular stomatitis virus spike glycoprotein (VSV-G) in human cells to induce gesicle release.
- Biochemical and functional analyses of gesicles to confirm protein incorporation and delivery capabilities.
- Delivery of TetR transactivator and murine leukemia virus (MLV) receptor (mCAT-1) using gesicles into various cell types.
- Assessment of gesicle-mediated mCAT-1 transfer for conferring permissiveness to ecotropic vectors in human fibroblasts.
Main Results:
- Engineered vesicles (gesicles) were successfully generated from human cells overexpressing VSV-G.
- Gesicles were shown to incorporate and deliver various proteins, including TetR and mCAT-1, to recipient cells.
- Gesicle-mediated transfer of mCAT-1 enabled the generation of human-induced pluripotent stem cells using ecotropic vectors in BSL-2 facilities.
- This method demonstrated enhanced biosafety for retro/lentiviral vector applications.
Conclusions:
- Gesicles represent a novel and versatile platform for the non-genetic delivery of proteins into human cells.
- This protein-transduction method offers significant potential for advancing research and therapeutic applications by enabling efficient and safe protein delivery.
- Gesicle technology holds promise for improving the safety of stem cell research and other cell-based applications involving viral vectors.
