Numerical operations in living cells by programmable RNA devices.

Kei Endo,Karin Hayashi,Hirohide Saito

doi:10.1126/sciadv.aax0835

Abstract

Integrated bioengineering systems can make executable decisions according to the cell state. To sense the state, multiple biomarkers are detected and processed via logic gates with synthetic biological devices. However, numerical operations have not been achieved. Here, we show a design principle for messenger RNA (mRNA) devices that recapitulates intracellular information by multivariate calculations in single living cells. On the basis of this principle and the collected profiles of multiple microRNA activities, we demonstrate that rationally programmed mRNA sets classify living human cells and track their change during differentiation. Our mRNA devices automatically perform multivariate calculation and function as a decision-maker in response to dynamic intracellular changes in living cells.

Highlights

To engineer living cells and organisms, artificial systems that function in response to cellular states have been programmed using synthetic devices made of biomolecules [1,2,3]
We designed synthetic messenger RNA (mRNA) that respond to multiple miRNA inputs and examined their behavior
The activity of an miRNA in a living cell can be efficiently detected by an mRNA that contains a completely complementary sequence to the target miRNA in the 5′ untranslated region (5′UTR) and expresses a reporter fluorescent protein [19, 20]

Summary

Introduction

To engineer living cells and organisms, artificial systems that function in response to cellular states have been programmed using synthetic devices made of biomolecules [1,2,3]. Switches [4,5,6,7,8,9,10], sometimes with inverters [11, 12], and split systems [13] sense a biomarker to change their activity, transmitting cellular information to the device. Logic gates [11, 14,15,16,17] process multiple inputs of biomarkers, extending the decision-making capacity. Molecular devices that perform multivariate numerical operations in living cells remain undeveloped. The number of applicable devices for sensing, signal transmitting, and decision-making limits the number of accessible biomarkers

Methods

Results

Conclusion