Paper-Based Synthetic Gene Networks

Abstract

Synthetic gene networks have wide-ranging uses in reprogramming and rewiring organisms. To date, there has not been a way to harness the vast potential of these networks beyond the constraints of a laboratory or in vivo environment. Here, we present an in vitro paper-based platform that provides a new venue for synthetic biologists to operate, and a much-needed medium for the safe deployment of engineered gene circuits beyond the lab. Commercially available cell-free systems are freeze-dried onto paper, enabling the inexpensive, sterile and abiotic distribution of synthetic biology-based technologies for the clinic, global health, industry, research and education. For field use, we create circuits with colorimetric outputs for detection by eye, and fabricate a low-cost, electronic optical interface. We demonstrate this technology with small molecule and RNA actuation of genetic switches, rapid prototyping of complex gene circuits, and programmable in vitro diagnostics, including glucose sensors and strain-specific Ebola virus sensors.

Portable, On-Demand Biomolecular Manufacturing

“…These limitations impact distribution costs and highlight the challenge of delivering the benefits of these technologies to developing regions. We recently reported a method for the safe deployment of genetically encoded tools (Pardee et al, 2014). Using freeze-dried, cell-free (FD-CF) expression machinery on paper, we generated a platform that retains the fundamental protein synthesis capability of live cells while remaining abiotic, sterile, and portable.…”

“…Using freeze-dried, cell-free (FD-CF) expression machinery on paper, we generated a platform that retains the fundamental protein synthesis capability of live cells while remaining abiotic, sterile, and portable. In combination with toehold switch RNA sensors (Green et al, 2014), this platform was used to demonstrate a new class of low-cost diagnostic tools (Pardee et al, 2016).…”

“…These advances have thus far been tied to laboratory settings where the necessary skills and equipment are found. Building off of this foundation, the proposed use of FD-CF systems, with their long-term activity at room temperature (> 1 year) and ease of operation, could alleviate both the restrictions of live-cell biosynthesis and cold-chain distribution requirements (Pardee et al, 2014). Recent reports draw emphasis to a pressing need for the decentralization of therapeutic biomanufacturing, offering novel alternatives that, nonetheless, require expensive, large equipment and highly skilled operators or yet rely on production from living cells (Adamo et al, 2016;Perez-Pinera et al, 2016).…”

Towards autonomous lab-on-a-chip devices for cell phone biosensing

“…Cell-free synthetic biology (CFSB (Smith et al, 2014;Pardee et al, 2014)) is a powerful bio-sensing concept that exploits the biochemical machinery in genetically engineered cells without the disadvantages of keeping the cells alive or dealing with issues of biosafety. Such a generic approach brings together the benefits of selectivity to pathogen-specific nucleotide sequences and signal amplification that can be co-opted from a model organism for a defined purpose.…”

Bioluminescent Antibodies for Point‐of‐Care Diagnostics

“…We therefore investigated whether the Fab‐based sensors would be compatible with measurements on paper. For this, we pretreated paper with bovine serum albumin and the non‐ionic detergent TWEEN 20 to reduce adsorption of the sensor and analytes, as well as to improve diffusion of the spotted liquid 23. The LUCIDs were then lyophilized on the pre‐treated paper for later use.…”