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In vitro incorporation of a cell‐binding protein to a lentiviral vector using an engineered split intein enables targeted delivery of genetic cargo
Author(s) -
ChamounEmanuelli Ana M.,
Wright Gus,
Roger Smith,
Münch Robert C.,
Buchholz Christian J.,
Chen Zhilei
Publication year - 2015
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.25685
Subject(s) - intein , homing (biology) , viral vector , fusion protein , gene delivery , computational biology , biology , in vitro , green fluorescent protein , protein engineering , microbiology and biotechnology , synthetic biology , lentivirus , genetic enhancement , gene , rna splicing , virology , genetics , biochemistry , virus , recombinant dna , ecology , rna , enzyme , viral disease
Gene therapy represents a promising therapeutic paradigm for addressing many disorders, but the absence of a vector that can be robustly and reproducibly functionalized with cell‐homing functionality to mediate the delivery of genetic cargo specifically to target cells following systemic administration has stood as a major impediment. In this study, a high‐affinity protein–protein pair comprising a splicing‐deficient naturally split intein was used as molecular Velcro to append a HER2/neu‐binding protein (DARPin) onto the surface of a binding‐deficient, fusion‐competent lentivirus. HER2/neu‐specific lentiviruses created using this in vitro pseudotyping approach were able to deliver their genetic reporter cargo specifically to cells that express the target receptor at high levels in a co‐culture. We envision that the described technology could provide a powerful, broadly applicable platform for the incorporation of cell‐targeting functionality onto viral vectors. Biotechnol. Bioeng. 2015;112: 2611–2617. © 2015 Wiley Periodicals, Inc.