Vectors

The TRON Vectors Unit is part of the Immunotherapy Development Center (IDC) and provides state-of-the-art gene transfer technologies to other units and teams of TRON. A key focus is on retroviral gene transfer. However we also expand our spectrum of methods on an ongoing basis by establishing aditional gene transfer techniques, such as transposons and alphaviral vectors.

Viruses have evolved specialized molecular mechanisms to efficiently transport their genomes inside cells, which make viral vectors ideal tools for delivering genetic material into cells. The delivery of genes can be performed inside a living organism (in vivo) or in cell culture (in vitro). Achieving uniformly efficient gene transfer to a wide variety of target cells is challenging. For this reason, we work with a modular set of tools by combining specialized vector backbones, various envelope proteins and optimized protocols.

The health risk associated with the natural viruses is eliminated by removing their pathologic potential from viral vectors and inhibiting their ability to spread from cell to cell: the virus are transformed into simple containers that can enter target cells and deliver the genes. To achieve this high level of biosafety, the viral genome is split. The genes required for building a functional viral particle are delivered with a so-called packaging plasmid, while the genetic information intended for delivery is located on a transfer vector.

At TRON we combine standard techniques such as transfection, electroporation and fluorescence activated cell sorting (FACS) with our own set of tools, including gamma-retroviral vectors, self-inactivating lentiviral vectors, shRNA and micro-RNA expression cassettes, PiggyBac transposons and Semliki forest virus derived alphaviral vectors.

To standardize procedures and accelerate the process of cloning genes of interest, all our transfer vectors have been designed as gateway® destination vectors. For shRNA or miRNA expression, the TRON Vectors Unit provides precut entry clones ready for ligation.

The production of retro- and lentiviral cell culture supernatants is routine in our unit. We ensure highly efficient transduction of target cells by producing viral supernatants with standardized protocols. If required, we also provide concentration and purification of the supernatants using PEG-precipitation protocols to yield high titer supernatants.

For the transduction of target cells we have established protocols that work with a wide variety of cells and also design customized protocols if requested. Feel free to contact us for more information.