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A synthetic mammalian electro-genetic transcription circuit.

Weber W, Luzi S, Karlsson M, Sanchez-Bustamante CD, Frey U, Hierlemann A, Fussenegger M - Nucleic Acids Res. (2009)

Bottom Line: Electric signal processing has evolved to manage rapid information transfer in neuronal networks and muscular contraction in multicellular organisms and controls the most sophisticated man-built devices.Using a synthetic biology approach to assemble electronic parts with genetic control units engineered into mammalian cells, we designed an electric power-adjustable transcription control circuit able to integrate the intensity of a direct current over time, to translate the amplitude or frequency of an alternating current into an adjustable genetic readout or to modulate the beating frequency of primary heart cells.Successful miniaturization of the electro-genetic devices may pave the way for the design of novel hybrid electro-genetic implants assembled from electronic and genetic parts.

View Article: PubMed Central - PubMed

Affiliation: ETH Zurich, Department of Biosystems Science and Engineering, Mattenstrasse 26, CH-4058 Basel, Switzerland.

ABSTRACT
Electric signal processing has evolved to manage rapid information transfer in neuronal networks and muscular contraction in multicellular organisms and controls the most sophisticated man-built devices. Using a synthetic biology approach to assemble electronic parts with genetic control units engineered into mammalian cells, we designed an electric power-adjustable transcription control circuit able to integrate the intensity of a direct current over time, to translate the amplitude or frequency of an alternating current into an adjustable genetic readout or to modulate the beating frequency of primary heart cells. Successful miniaturization of the electro-genetic devices may pave the way for the design of novel hybrid electro-genetic implants assembled from electronic and genetic parts.

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Mammalian cell-based frequency generator. (a) Circuit diagram of the cell-based frequency generator. DC power converts ethanol into acetaldehyde, which dose-dependently triggers expression of the BMP-2 in engineered rat cardiomyocytes (AIRNRC-BMP-2) and increases the contraction frequency (tachycardia). (b) The beating frequency of cardiomyocytes is recorded as a function of the input current and acetaldehyde concentration using a CMOS-based HD-MEA. NC: negative control, mock-transduced cardiomyocytes; PC: positive control, cells transduced for constitutive BMP-2 expression. (c) HD-MEA-based analysis of the electrogenic behaviour of NRCs engineered for electro-inducible acetaldehyde-responsive BMP-2 expression. The dataset shown as example was recorded at a direct input current of 50 mA corresponding to a beating frequency of 2.1 Hz. Detailed activation map illustrating the average signal shape of the 121 selected electrodes during 10 s. Average signal shape over all 121 channels. Raster plot showing a dot for each contraction on each channel over time. Inter-burst interval or inter-beat interval used to calculate the average beating frequency and beating frequency variation. (d) Zoom-in of two selected bursts/beats on different electrodes. (e) Long signal trace showing the synchronized contraction frequency on five selected electrodes or channels.
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Figure 3: Mammalian cell-based frequency generator. (a) Circuit diagram of the cell-based frequency generator. DC power converts ethanol into acetaldehyde, which dose-dependently triggers expression of the BMP-2 in engineered rat cardiomyocytes (AIRNRC-BMP-2) and increases the contraction frequency (tachycardia). (b) The beating frequency of cardiomyocytes is recorded as a function of the input current and acetaldehyde concentration using a CMOS-based HD-MEA. NC: negative control, mock-transduced cardiomyocytes; PC: positive control, cells transduced for constitutive BMP-2 expression. (c) HD-MEA-based analysis of the electrogenic behaviour of NRCs engineered for electro-inducible acetaldehyde-responsive BMP-2 expression. The dataset shown as example was recorded at a direct input current of 50 mA corresponding to a beating frequency of 2.1 Hz. Detailed activation map illustrating the average signal shape of the 121 selected electrodes during 10 s. Average signal shape over all 121 channels. Raster plot showing a dot for each contraction on each channel over time. Inter-burst interval or inter-beat interval used to calculate the average beating frequency and beating frequency variation. (d) Zoom-in of two selected bursts/beats on different electrodes. (e) Long signal trace showing the synchronized contraction frequency on five selected electrodes or channels.

Mentions: Electric signals linked to complex intracellular signalling cascades are well-known to manage muscular contraction in specialized mammalian cells (38). For example, cardiac ventricular contraction frequency of NRCs is modulated by BMP-2 that induces receptor-mediated activation of the myocyte-specific enhancer factor 2A via phosphatidylinositol 3-kinase in a dose-dependent manner (39). By transducing NRCs cultivated on complementary metal-oxide semiconductor (CMOS)-based HD-MEAs with lentiviral particles (32) engineered for electro-inducible acetaldehyde-responsive expression of BMP-2, we were able to convert DC into an oscillating electronic signal with a defined frequency (Figure 3a). Challenging this cell-based frequency generator with increasing input current resulted in elevated BMP-2 expression which stimulated NRCs to beat at higher frequency as scored by the HD-MEA (Figure 3b–e).Figure 3.


A synthetic mammalian electro-genetic transcription circuit.

Weber W, Luzi S, Karlsson M, Sanchez-Bustamante CD, Frey U, Hierlemann A, Fussenegger M - Nucleic Acids Res. (2009)

Mammalian cell-based frequency generator. (a) Circuit diagram of the cell-based frequency generator. DC power converts ethanol into acetaldehyde, which dose-dependently triggers expression of the BMP-2 in engineered rat cardiomyocytes (AIRNRC-BMP-2) and increases the contraction frequency (tachycardia). (b) The beating frequency of cardiomyocytes is recorded as a function of the input current and acetaldehyde concentration using a CMOS-based HD-MEA. NC: negative control, mock-transduced cardiomyocytes; PC: positive control, cells transduced for constitutive BMP-2 expression. (c) HD-MEA-based analysis of the electrogenic behaviour of NRCs engineered for electro-inducible acetaldehyde-responsive BMP-2 expression. The dataset shown as example was recorded at a direct input current of 50 mA corresponding to a beating frequency of 2.1 Hz. Detailed activation map illustrating the average signal shape of the 121 selected electrodes during 10 s. Average signal shape over all 121 channels. Raster plot showing a dot for each contraction on each channel over time. Inter-burst interval or inter-beat interval used to calculate the average beating frequency and beating frequency variation. (d) Zoom-in of two selected bursts/beats on different electrodes. (e) Long signal trace showing the synchronized contraction frequency on five selected electrodes or channels.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2651811&req=5

Figure 3: Mammalian cell-based frequency generator. (a) Circuit diagram of the cell-based frequency generator. DC power converts ethanol into acetaldehyde, which dose-dependently triggers expression of the BMP-2 in engineered rat cardiomyocytes (AIRNRC-BMP-2) and increases the contraction frequency (tachycardia). (b) The beating frequency of cardiomyocytes is recorded as a function of the input current and acetaldehyde concentration using a CMOS-based HD-MEA. NC: negative control, mock-transduced cardiomyocytes; PC: positive control, cells transduced for constitutive BMP-2 expression. (c) HD-MEA-based analysis of the electrogenic behaviour of NRCs engineered for electro-inducible acetaldehyde-responsive BMP-2 expression. The dataset shown as example was recorded at a direct input current of 50 mA corresponding to a beating frequency of 2.1 Hz. Detailed activation map illustrating the average signal shape of the 121 selected electrodes during 10 s. Average signal shape over all 121 channels. Raster plot showing a dot for each contraction on each channel over time. Inter-burst interval or inter-beat interval used to calculate the average beating frequency and beating frequency variation. (d) Zoom-in of two selected bursts/beats on different electrodes. (e) Long signal trace showing the synchronized contraction frequency on five selected electrodes or channels.
Mentions: Electric signals linked to complex intracellular signalling cascades are well-known to manage muscular contraction in specialized mammalian cells (38). For example, cardiac ventricular contraction frequency of NRCs is modulated by BMP-2 that induces receptor-mediated activation of the myocyte-specific enhancer factor 2A via phosphatidylinositol 3-kinase in a dose-dependent manner (39). By transducing NRCs cultivated on complementary metal-oxide semiconductor (CMOS)-based HD-MEAs with lentiviral particles (32) engineered for electro-inducible acetaldehyde-responsive expression of BMP-2, we were able to convert DC into an oscillating electronic signal with a defined frequency (Figure 3a). Challenging this cell-based frequency generator with increasing input current resulted in elevated BMP-2 expression which stimulated NRCs to beat at higher frequency as scored by the HD-MEA (Figure 3b–e).Figure 3.

Bottom Line: Electric signal processing has evolved to manage rapid information transfer in neuronal networks and muscular contraction in multicellular organisms and controls the most sophisticated man-built devices.Using a synthetic biology approach to assemble electronic parts with genetic control units engineered into mammalian cells, we designed an electric power-adjustable transcription control circuit able to integrate the intensity of a direct current over time, to translate the amplitude or frequency of an alternating current into an adjustable genetic readout or to modulate the beating frequency of primary heart cells.Successful miniaturization of the electro-genetic devices may pave the way for the design of novel hybrid electro-genetic implants assembled from electronic and genetic parts.

View Article: PubMed Central - PubMed

Affiliation: ETH Zurich, Department of Biosystems Science and Engineering, Mattenstrasse 26, CH-4058 Basel, Switzerland.

ABSTRACT
Electric signal processing has evolved to manage rapid information transfer in neuronal networks and muscular contraction in multicellular organisms and controls the most sophisticated man-built devices. Using a synthetic biology approach to assemble electronic parts with genetic control units engineered into mammalian cells, we designed an electric power-adjustable transcription control circuit able to integrate the intensity of a direct current over time, to translate the amplitude or frequency of an alternating current into an adjustable genetic readout or to modulate the beating frequency of primary heart cells. Successful miniaturization of the electro-genetic devices may pave the way for the design of novel hybrid electro-genetic implants assembled from electronic and genetic parts.

Show MeSH
Related in: MedlinePlus