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DNA watermarks in non-coding regulatory sequences.

Heider D, Pyka M, Barnekow A - BMC Res Notes (2009)

Bottom Line: In a third approach we introduced a second overlapping watermark into the lac promoter, which did not affect the promoter activity.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 accordingly.Therefore, we do not recommend integrating watermark sequences into regulatory regions.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Experimental Tumorbiology, University of Münster, Badestr, 9, D-48149 Münster, Germany. dominik.heider@uni-due.de

ABSTRACT

Background: DNA 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.

Findings: The 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.

Conclusion: 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 accordingly. Therefore, we do not recommend integrating watermark sequences into regulatory regions.

No MeSH data available.


Related in: MedlinePlus

Growth characteristics. Growth characteristics of strain YRA130 and the Yep352-SCR1 and Yep352-SCR1-TB transformed YRA130 strains in YPD full medium (A) and SD-ura medium (B). Three independent experiments with a standard deviation <0.03 are shown. All data show p-values <0.05, except YRA130 cells transformed with Yep352-SCR1 compared with YRA130 cells transformed with Yep352-SCR1-TB.
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Figure 2: Growth characteristics. Growth characteristics of strain YRA130 and the Yep352-SCR1 and Yep352-SCR1-TB transformed YRA130 strains in YPD full medium (A) and SD-ura medium (B). Three independent experiments with a standard deviation <0.03 are shown. All data show p-values <0.05, except YRA130 cells transformed with Yep352-SCR1 compared with YRA130 cells transformed with Yep352-SCR1-TB.

Mentions: Although the predicted secondary structure of the watermarked scR1 differs from the wild type structure, the resulting RNA molecule proved to be functional active indirectly demonstrated by the YRA130 cells transformed either with the Yep352-SCR1 or the Yep352-SCR1-TB, which displayed no significant differences in their growth characteristics (Figure 2).


DNA watermarks in non-coding regulatory sequences.

Heider D, Pyka M, Barnekow A - BMC Res Notes (2009)

Growth characteristics. Growth characteristics of strain YRA130 and the Yep352-SCR1 and Yep352-SCR1-TB transformed YRA130 strains in YPD full medium (A) and SD-ura medium (B). Three independent experiments with a standard deviation <0.03 are shown. All data show p-values <0.05, except YRA130 cells transformed with Yep352-SCR1 compared with YRA130 cells transformed with Yep352-SCR1-TB.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2713970&req=5

Figure 2: Growth characteristics. Growth characteristics of strain YRA130 and the Yep352-SCR1 and Yep352-SCR1-TB transformed YRA130 strains in YPD full medium (A) and SD-ura medium (B). Three independent experiments with a standard deviation <0.03 are shown. All data show p-values <0.05, except YRA130 cells transformed with Yep352-SCR1 compared with YRA130 cells transformed with Yep352-SCR1-TB.
Mentions: Although the predicted secondary structure of the watermarked scR1 differs from the wild type structure, the resulting RNA molecule proved to be functional active indirectly demonstrated by the YRA130 cells transformed either with the Yep352-SCR1 or the Yep352-SCR1-TB, which displayed no significant differences in their growth characteristics (Figure 2).

Bottom Line: In a third approach we introduced a second overlapping watermark into the lac promoter, which did not affect the promoter activity.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 accordingly.Therefore, we do not recommend integrating watermark sequences into regulatory regions.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Experimental Tumorbiology, University of Münster, Badestr, 9, D-48149 Münster, Germany. dominik.heider@uni-due.de

ABSTRACT

Background: DNA 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.

Findings: The 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.

Conclusion: 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 accordingly. Therefore, we do not recommend integrating watermark sequences into regulatory regions.

No MeSH data available.


Related in: MedlinePlus