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Engineering Cell-permeable Protein
Published on: December 28, 2009
Efficient CPP-mediated Cre protein delivery to developing and adult CNS tissues.
Yorick Gitton1, Lorenzo Tibaldi, Edmond Dupont
1Evolution des Régulations Endocriniennes, CNRS UMR 7221, Muséum National d'Histoire Naturelle, 7 rue Cuvier,75005 Paris, France. gitton@mnhn.fr
BMC Biotechnology
|April 28, 2009
Summary
Direct protein delivery using Cre recombinase conjugates offers a novel method for targeted genetic modification in central nervous system tissues. This approach enables precise control over gene function studies and modeling of neurodegenerative processes.
Area of Science:
- Neuroscience
- Molecular Biology
- Genetics
Background:
- Gene function manipulation is crucial for fundamental and applied research.
- Current methods rely on genetic or viral/non-viral gene delivery strategies.
- This study introduces direct protein delivery to central nervous system (CNS) tissues.
Purpose of the Study:
- To develop and evaluate a direct protein delivery strategy for genetic modification in CNS tissues.
- To assess the efficacy of Cre recombinase-cell-penetrating peptide conjugates in neural explants.
- To establish tools for spatially restricted genetic modification and study of neural cell dynamics.
Main Methods:
- Fusion of Cre recombinase with cell-penetrating peptides to create active protein conjugates.
- Delivery of Cre conjugates to adherent cultured cells and organotypic neural tissue explants.
- Comparison of direct protein addition and conjugate-coated bead implantation methods.
Main Results:
- Cre recombinase conjugates were internalized and active in cell nuclei.
- Genomic recombination was achieved in neural explants via both delivery methods.
- Bead implantation allowed for spatially restricted recombination in neural tissues.
- Delivery to adult brain tissue transduced mature, postmitotic neurons.
Conclusions:
- Direct protein delivery provides tools for spatially restricted genetic modification in explant cultures.
- This strategy facilitates studies on neural cell lineage, migration, differentiation, and death.
- Demonstrated selective elimination of spinal cord interneurons for neurodegenerative process modeling.
- Opens new avenues for cell targeting protocols using bioactive proteic cargoes.
