Intracellular trafficking of a dynein-based nanoparticle designed for gene delivery

Author(s)

de Pinho Favaro, Marianna Teixeira ^{[1, 2]} ; Unzueta, Ugutz ^{[3, 4]} ; de Cabo, Marti ^[5] ; Villaverde, Antonio ^{[2, 3, 6]} ; Ferrer-Miralles, Neus ^{[2, 3, 6]} ; Azzoni, Adriano Rodrigues ^[7]

Affiliation

^[1] Univ Estadual Campinas, Ctr Biol Mol & Engn Genet, Av Candido Rondon 400, BR-13083875 Campinas, SP - Brazil

^[2] Univ Autonoma Barcelona, Inst Biotecnol & Biomed, E-08193 Barcelona - Spain

^[3] CIBER Bioingn Biomat & Nanomed CIBER BBN, Barcelona 08193 - Spain

^[4] Hosp Santa Creu & Sant Pau, Inst Invest Biomed St Pau, Barcelona 08025 - Spain

^[5] Univ Autonoma Barcelona, Serv Microscopia, E-08193 Barcelona - Spain

^[6] Univ Autonoma Barcelona, Dept Genet & Microbiol, E-08193 Barcelona - Spain

^[7] Univ Sao Paulo, Escola Politecn, Dept Engn Quim, Av Prof Luciano Gualberto, Trav 3, 380, BR-05508900 Sao Paulo, SP - Brazil

• Document type: Journal article
• Source: European Journal of Pharmaceutical Sciences; v. 112, p. 71-78, JAN 15 2018.
• Web of Science Citations: 1

Abstract

The success of viruses in the delivery of the viral genome to target cells relies on the evolutionary selection of protein-based domains able to hijack the intermolecular interactions through which cells respond to intra-and extracellular stimuli. In an effort to mimic viral infection capabilities during non-viral gene delivery, a modular recombinant protein named T-Rp3 was recently developed, containing a DNA binding domain, a dynein molecular motor interacting domain, and a TAT-derived transduction domain. Here, we analyzed at the microscopic level the mechanisms behind the cell internalization and intracellular trafficking of this highly efficient modular protein vector. We found that the protein has the ability to self-assemble in discrete protein nanoparticles resembling viral capsids, to bind and condense plasmid DNA (pDNA), and to interact with eukaryotic cell membranes. Confocal and single particle tracking assays performed on living HeLa cells revealed that the T-Rp3 nanoparticles promoted an impressive speed of cellular uptake and perinuclear accumulation. Finally, the protein demonstrated to be a versatile vector, delivering siRNA at efficiencies comparable to Lipofectamine T. These results demonstrate the high potential of recombinant modular proteins with merging biological functions to fulfill several requirements needed to obtain cost-effective non-viral vectors for gene-based therapies. (AU)

• FAPESP's process: 12/18850-8 - Development and in vitro and in vivo evaluation of gene delivery vectors based on dynein light chain Rp3 and synthetic peptides.
• Grantee: Marianna Teixeira de Pinho Favaro
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 13/23780-1 - Development of non-viral gene delivery vectors based on the Rp3 Dynein light chain
• Grantee: Adriano Rodrigues Azzoni
• Support Opportunities: Regular Research Grants