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Paper-based Synthetic Gene Networks

A technology for synthesizing genes and networks, which is applied in the field of paper-based synthetic gene networks, and can solve problems such as limiting equipment applications

Active Publication Date: 2022-04-05
PRESIDENT & FELLOWS OF HARVARD COLLEGE +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, both living cells and synthetic liposomes suffer from severe drawbacks in terms of practical issues such as storage and transport, thereby limiting the configuration of devices based on synthetic gene networks and their applications.
In particular, it is not possible to use these devices for on-demand applications

Method used

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  • Paper-based Synthetic Gene Networks
  • Paper-based Synthetic Gene Networks
  • Paper-based Synthetic Gene Networks

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0365] Example 1: Lyophilized sensors comprising synthetic gene networks and cell-free systems

[0366] showed for the first time that freeze-dried cell extracts can support constitutive expression of GFP via Escherichia coli RNA polymerase or T7 RNA polymerase-based promoters ( Figure 1A ). Subsequent T7-based expression can also be supported by lyophilization of recombinant protein-based systems. These results essentially reflect the expression levels observed in fresh cell-free reactions and in E. coli. The lyophilized cell-free system is stable over time, with transcription and translation remaining highly active after several months of storage at room temperature ( Figure 1B ).

[0367] Inducible expression systems are used to synthesize gene networks. GFP expression is readily induced by the antibiotic doxycycline or chemical analogues using the classic inducible tetO promoter. Notably, tight regulatory control in vitro requires recruitment of the tet repressor (t...

Embodiment 2

[0373] Example 2: Paper-based Synthetic Gene Networks

[0374] Bacterial and mammalian assemblies can be freeze-dried on paper and other porous substrates to create quiescent synthetic gene networks that are stable for long-term storage at room temperature and can be activated by rehydration. The resulting engineered material has the transcriptional and translational properties of the cell and can accommodate genetically encoded tools using, for example, commercially available cell-free transcription and translation systems. Shown here are techniques for RNA actuation with small molecules and gene switches, construction of paper-based sensors for glucose and mRNA (including antibiotic resistance genes), and characterization of new genetic circuits. For greater practical use, genetic circuits could be augmented with colorimetric output (for detection by the naked eye), and by low-cost fabrication, an electro-optical interface for quantification and possibly automated reactions....

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Abstract

Disclosed herein are shelf-stable compositions based on synthetic gene networks and / or cell-free systems that are lyophilized on solid supports. The composition can be easily transported and stored for a period of time and can be activated by simply adding water. Methods of use are also disclosed herein, including but not limited to sensing and various logic functions. The present invention enables simple, sterile and abiotic deployment of synthetic biology-based technologies into clinical settings, food processing and industrial, military and consumer products.

Description

[0001] Cross References to Related Applications [0002] Pursuant to 35 U.S.C. §119(e), this application claims the benefit of U.S. Provisional Application No. 61 / 913,110, filed December 6, 2013, and U.S. Provisional Application No. 62 / 066,966, filed October 22, 2014, to The contents of each are incorporated herein in their entirety by reference. technical field [0003] The present disclosure generally relates to shelf-stable compositions and methods for sensing and performing synthetic gene network-based logic functions. Background technique [0004] The field of synthetic biology aims to redesign components of natural systems to solve global challenges, such as reducing the cost of drug production or green energy and chemistry. In doing so, synthetic biologists are creating whole-cell biosensors, synthetic probiotics, and new sources of medicine and green energy and novel chemistry. At the heart of these technologies is a network of synthetic genes that control the cell...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6806
CPCC12Q1/6806C12Q2521/119C12Q2527/125C12Q1/6897
Inventor 凯特·帕迪詹姆斯·J·柯林斯
Owner PRESIDENT & FELLOWS OF HARVARD COLLEGE