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Author
Sangés Ametllé, Marina
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Abstract
In modern biotechnology and DNA bioengineering there is a high demand of new processes that allow quick and efficient DNA modifications. The use of large-serine recombinases (LSRs) is an innovative tool for advancing synthetic biology. The controlled rearrangement and modification of DNA in gene therapy, biotechnology and synthetic biology of the LSRs has become one of the most attractive characteristics for upcoming investigations.
For the use of LSRs it is important to have a pure cell line with the att landing pads specific for each recombinase in order to let the recombinase recognise the specific zone of the genome and recombine the genes trough Recombinase-Mediated Cassette Exchange (RMCE) or the insertion of the whole plasmid.
In this thesis, we will follow all the steps necessary to create a pure cell line with the attP landing pad of Bxb1 recombinase including all the cloning steps and the confirmation that the landing pad is successfully inserted inside the cell. On the other hand, we will test the efficiency of Cre recombinase for the recombination trough RMCE with the antibody Herceptin heavy and light chain.
This project is contemplating all the necessary steps to modify a wild type CHO cell with landing pads for a specific recombinase and test the efficiency of the recombinase. Future investigations will test different recombinases following the same process to optimise the recombination process and allow synthetic biology progress.
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