title: Improved performance of protein-based recombinant gene therapy vehicles by tuning downstream procedures.
abstract: Protein engineering offers a robust platform for the design and production in cell factories of a plethora of protein-based drugs, including nonviral gene therapy vehicles. We have determined here that a protein nanoparticle, formed by highly cationic protein monomers, fails to bind exogenous DNA and to promote detectable gene expression in target cells despite recruiting all the needed functions. Removal of DNA and RNA with nucleases previous to forming complexes with exogenous DNA dramatically enhances the ability of the protein to bind and transfer DNA to target cell nuclei. These data point out contaminant nucleic acids deriving from the cell factory as a major factor impairing the performance of protein-based artificial viruses and stress the need of a nuclease step in the downstream of proteins whose function is based on cationic domains.
authors: Institut de Biotecnologia i de Biomedicina, Universitat Autònoma de Barcelona, Bellaterra, 08193, Barcelona, Spain; CIBER de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN), Bellaterra, 08193, Barcelona, Spain; Department de Genètica i de Microbiologia, Universitat Autònoma de Barcelona, Bellaterra, 08193, Barcelona, Spain.
authors: Ugutz
authors: U
authors: Unzueta
authors: Paolo
authors: P
authors: Saccardo
authors: Neus
authors: N
authors: Ferrer-Miralles
authors: Elena
authors: E
authors: García-Fruitós
authors: Esther
authors: E
authors: Vazquez
authors: Antonio
authors: A
authors: Villaverde
authors: Francisco
authors: F
authors: Cortés
authors: Ramón
authors: R
authors: Mangues
mesh_terms: D004247:DNA / Q000737:chemistry / Q000235:genetics; D015870:Gene Expression; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D053758:Nanoparticles / Q000737:chemistry*; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics*
chemical_list: D011994:Recombinant Proteins; D004247:DNA
keywords: downstream; gene therapy; nanoparticles; nucleic acids; recombinant proteins