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Molecular breeding of viruses

Nature Genetics volume 25, pages 436–439 (2000)Cite this article

Abstract

Genetic recombination is a major force driving the evolution of many viruses. Recombination between two copackaged retroviral genomes may occur at rates as high as 40% per replication cycle1. This enables genetic information to be shuffled rapidly, leading to recombinants with new patterns of mutations and phenotypes. The in vitro process of DNA shuffling2,3 (molecular breeding) mimics this mechanism on a vastly parallel and accelerated scale. Multiple homologous parental sequences are recombined in parallel, leading to a diverse library of complex recombinants from which desired improvements can be selected. Different proteins and enzymes have been improved using DNA shuffling4,5,6. We report here the first application of molecular breeding to viruses. A single round of shuffling envelope sequences from six murine leukaemia viruses (MLV) followed by selection yielded a chimaeric clone with a completely new tropism for Chinese Hamster Ovary (CHOK1) cells. The composition and properties of the selected clone indicated that this particular permutation of parental sequences cannot be readily attained by natural retroviral recombination. This example demonstrates that molecular breeding can enhance the inherently high evolutionary potential of retroviruses to obtain desired phenotypes. It can be an effective tool, when information is limited, to optimize viruses for gene therapy and vaccine applications when multiple complex functions must be simultaneously balanced.

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Figure 1: Construction of shuffled MLV envelope library.
Figure 2: Structure of the evolved chimaeric envelope showing segments contributed by the three Friend parents.
Figure 3: Distinct tropisms of parental and chimaeric viruses.

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Acknowledgements

We thank J. Minshull, R. Shibata, P. Patten, O. Danos, F. Fan and J.Y. Han for useful input; S. Goff for the pNCA Moloney molecular clone; and Gene Therapy Laboratories for the G1 retroviral vector.

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  1. Maxygen Inc., Redwood City, California , USA

    Nay-Wei Soong, Laurel Nomura, Katja Pekrun, Margaret Reed, Liana Sheppard, Glenn Dawes & Willem P.C. Stemmer

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  1. Nay-Wei Soong
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Correspondence to Nay-Wei Soong.

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Soong, NW., Nomura, L., Pekrun, K. et al. Molecular breeding of viruses. Nat Genet 25, 436–439 (2000). https://doi.org/10.1038/78132

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