With the growing availability of low-cost de novo gene synthesis, synthetic biology not only allows unrestricted and flexible design of non-natural DNA sequences, but also adapting coding sequences to the genetic requirements of the chosen target organism. It promises combining technical engineering approaches with biological sciences and informatics to predict, simulate, and construct novel pathways, genomes and organisms faster and more precisely. The present dawn of synthetic biology opens up entirely new horizons in genetic engineering.
Until recently almost all genetic templates originated from natural sources, limiting the range of possibilities. Initially the focus was removing cross-species boundaries, rearranging natural genetic building blocks and introducing minor modifications into DNA sequences. This was transformed when modern molecular biology enabled systematic genetic manipulation and redesign of novel strains and genetically modified organisms. This does not alter the authors′ adherence to all the PLoS ONE policies on sharing data and materials, as detailed online in the guide for authors.įor millennia, mankind has employed the principle of consecutively selecting random mutations to breed desired phenotypes into crops, livestock, pets and microbes, relying on a trial-and-error approach. The authors also declare competing interests in the form of patent family US2011119778 including all national patents and pending applications. Wagner are employees of Life Technologies/Geneart AG, who performed gene design, optimization and synthesis. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Ĭompeting interests: The authors have read the journal’s policy and have the following conflicts: Dr. No additional external funding received for this study. Tobacco leaf transfection and analysis was funded by the Deutsche Forschungsgemeinschaft, grant HA3468/4-1. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.įunding: Assessment of HIV-1 infectivity and long-term replication kinetics was funded by the Bundesministerium fuer Bildung und Forschung, grant 0313687. Received: ApAccepted: JPublished: August 8, 2012Ĭopyright: © Liss et al. Tata Institute of Fundamental Research, India (2012) Embedding Permanent Watermarks in Synthetic Genes. We also introduced Vigenère polyalphabetic substitution to cipher text messages, and developed an identifier as a key to deciphering codon usage ranking stored for a specific organism within a sequence of 35 nucleotides.Ĭitation: Liss M, Daubert D, Brunner K, Kliche K, Hammes U, Leiherer A, et al. Several different messages were embedded into open reading frames of T7 RNA polymerase, GFP, human EMG1 and HIV gag, variously optimized for bacterial, yeast, mammalian or plant expression, without affecting their protein expression or function. Preferably amino acids with 4 or 6 synonymous codons are used to comprise binary digits. To aim for good expression of the labeled gene in its host as well as retain a high degree of codon assignment flexibility for gene optimization, codon usage tables of the target organism are taken into account.
Text messages are first translated into a binary string, and then represented in the reading frame by synonymous codon choice. We describe a method for embedding messages within open reading frames of synthetic genes by adapting steganographic algorithms typically used for watermarking digital media files. Ideally information should be inseparably interlaced into expressed genes. As synthetic biology advances, labeling of genes or organisms, like other high-value products, will become important not only to pinpoint their identity, origin, or spread, but also for intellectual property, classification, bio-security or legal reasons.
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