In vitro incorporation of a cell-binding protein to a lentiviral vector using an engineered split intein enables targeted delivery of genetic cargo

• Published in: Biotechnology and bioengineering Vol. 112; no. 12; pp. 2611 - 2617
• Main Authors: Chamoun-Emanuelli, Ana M., Wright, Gus, Roger III, Smith, Münch, Robert C., Buchholz, Christian J., Chen, Zhilei
• Format: Journal Article
• Language: English
• Published: United States Blackwell Publishing Ltd 01.12.2015; Wiley Subscription Services, Inc
• Subjects: Binding sites; Cargo; Carrier Proteins - genetics; Carrier Proteins - metabolism; Cell culture; Drug Delivery Systems; Gene therapy; Genetic engineering; Genetic Therapy - methods; Genetics; HER2/neu; In vitro testing; Lentivirus; Lentivirus - genetics; Lentivirus - physiology; Mathematical analysis; Platforms; Proteins; Receptor, ErbB-2 - metabolism; Receptors, Virus - metabolism; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Vectors (mathematics); Virus Attachment; gene therapy; HER2/neu; lentivirus
• ISSN: 0006-3592; 1097-0290
• DOI: 10.1002/bit.25685

ABSTRACT 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. A novel strategy for reprogramming the cell‐type specificity of lentiviral vectors via robust and reproducible in vitro attachment of a target‐cell‐binding protein to the lentivirus surface is described. A high‐affinity protein–protein pair comprising a splicing‐deficient naturally split intein is used to load a binding‐deficient, fusion‐competent lentivirus with a target‐cell‐binding molecule (“binder”). This approach could provide a powerful, broadly applicable platform for altering the cell‐type specificity of viral vectors for gene therapy applications.