The emerging age of cell‐free synthetic biology

Abstract

a b s t r a c tThe engineering of and mastery over biological parts has catalyzed the emergence of synthetic biology. This field has grown exponentially in the past decade. As increasingly more applications of synthetic biology are pursued, more challenges are encountered, such as delivering genetic material into cells and optimizing genetic circuits in vivo. An in vitro or cell-free approach to synthetic biology simplifies and avoids many of the pitfalls of in vivo synthetic biology. In this review, we describe some of the innate features that make cell-free systems compelling platforms for synthetic biology and discuss emerging improvements of cell-free technologies. We also select and highlight recent and emerging applications of cell-free synthetic biology.

Elucidating the potential of crude cell extracts for producing pyruvate from glucose

“…Unfortunately, efforts to engineer these systems to unlock their diverse metabolic potential require developing clever methodologies to overcome aspects of the machinery that the organism uses for survival. In vitro synthetic biology offers an alternative way to harness an organism's rich metabolism; it is driven by the prospect of easy to manipulate, static systems (1). Livings cells require membranes, energy and building blocks for growth and dynamic regulation of their biochemical processes.…”

Synthetic biology platform technologies for antimicrobial applications

“…In vitro cell free expression systems that contain transcription and translation machinery provide the environment necessary for biologically-based technologies to function independently of living cells [122]. Originally utilized for in vitro protein expression, cell free expression systems have recently been recognized for their ability to support more complicated gene networks [123][124][125][126].…”

“…Further, the more precise control of conditions offered by in vitro expression systems has enabled the high-throughput tuning of synthetic constructs [123,124,127,128]. Despite these advancements, the application of cell-free methods has been limited to laboratory settings due to strict storage requirements, including the need for refrigeration.…”

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.…”