Abstract
BackgroundDNA watermarks can be applied to identify the unauthorized use of genetically modified organisms. It has been shown that coding regions can be used to encrypt information into living organisms by using the DNA-Crypt algorithm. Yet, if the sequence of interest presents a non-coding DNA sequence, either the function of a resulting functional RNA molecule or a regulatory sequence, such as a promoter, could be affected. For our studies we used the small cytoplasmic RNA 1 in yeast and the lac promoter region of Escherichia coli.FindingsThe lac promoter was deactivated by the integrated watermark. In addition, the RNA molecules displayed altered configurations after introducing a watermark, but surprisingly were functionally intact, which has been verified by analyzing the growth characteristics of both wild type and watermarked scR1 transformed yeast cells. In a third approach we introduced a second overlapping watermark into the lac promoter, which did not affect the promoter activity.ConclusionEven though the watermarked RNA and one of the watermarked promoters did not show any significant differences compared to the wild type RNA and wild type promoter region, respectively, it cannot be generalized that other RNA molecules or regulatory sequences behave accordingly. Therefore, we do not recommend integrating watermark sequences into regulatory regions.
Highlights
DNA watermarks can be applied to identify the unauthorized use of genetically modified organisms
Even though the watermarked RNA and one of the watermarked promoters did not show any significant differences compared to the wild type RNA and wild type promoter region, respectively, it cannot be generalized that other RNA molecules or regulatory sequences behave
The lac operon of Escherichia coli consists of a promoter, three operators and three genes, coding for the β-galactoside permease, β-galactosidase and β-galactoside transacetylase, which are required for the lactose metabolism in Escherichia coli [5]
Summary
DNA watermarks can be applied to identify the unauthorized use of genetically modified organisms. DNA watermarks can be used for authenticating genetically modified organisms or in future for labeling animals in breeding [1,2]. It has already been shown in silico and in vivo that these watermarks do not affect the translation of proteins [1,2,3,4]. These assumptions only apply to coding regions, the insertion of watermarks into regulatory sequences like promoter regions or regulatory RNA molecules, had to be examined. The promoter sequence of the lac operon consists of two highly (page number not for citation purposes)
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