Abstract
Simple SummaryPromoters are DNA sequences where the process of transcription starts. They can work constitutively or be controlled by environmental signals of different types. The quantity of proteins and RNA present in yeast genetic circuits highly depends on promoter strength. Hence, they have been deeply studied and modified over, at least, the last forty years, especially since the year 2000 when Synthetic Biology was born. Here, we present how promoter engineering changed over these four decades and discuss its possible future directions due to novel computational methods and technology.Synthetic gene circuits are made of DNA sequences, referred to as transcription units, that communicate by exchanging proteins or RNA molecules. Proteins are, mostly, transcription factors that bind promoter sequences to modulate the expression of other molecules. Promoters are, therefore, key components in genetic circuits. In this review, we focus our attention on the construction of artificial promoters for the yeast S. cerevisiae, a popular chassis for gene circuits. We describe the initial techniques and achievements in promoter engineering that predated the start of the Synthetic Biology epoch of about 20 years. We present the main applications of synthetic promoters built via different methods and discuss the latest innovations in the wet-lab engineering of novel promoter sequences.
Highlights
Synthetic biology is a new branch of biology—whose birth can be set to January 2000 with the publication of the first two synthetic gene circuits [1,2]—that aims to standardize and modularize the design and engineering of biological circuits that confer new, useful functions to living cells [3].Among other applications, genetic circuits are constructed to produce high-value compounds, such as drugs, on an industrial scale [4,5]
We describe yeast synthetic promoters regulated by eight different kinds of bacterial proteins
Each Xylose-responsive transcription factors (XylRs) targeted a synthetic promoter made of the upstream activating sequence (UAS) from pTEF1 and the core GAL1 promoter was modified with a xyl operator downstream of the TATA box
Summary
Synthetic biology is a new branch of biology—whose birth can be set to January 2000 with the publication of the first two synthetic gene circuits [1,2]—that aims to standardize and modularize the design and engineering of biological circuits that confer new, useful functions to living cells [3]. The structure of a promoter is specified by the transcription start site (TSS) and the position, relative to the TSS, of various protein-binding sequences. S. cerevisiae RNA polymerase II-dependent promoters are characterized by three main elements: the upstream activating sequence (UAS), the TATA box, and the transcription start site [15]. Each of these structural motifs can be present in more than a single instance along the same promoter sequence. New synthetic promoters were designed by starting from two templates containing minimal motifs Each protein can be controlled by an environmental signal such that gene expression is induced or inhibited by either adding/removing chemicals to/from the cell solution or growing cells under particular conditions
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