![]() ![]() Experimental characterization of single or collections of natural or synthetic parts has also facilitated predictive models to assist with synthetic cistron construction ( 5, 11, 12).Įngineered genetic circuits consisting of one or a few genes have been successfully applied to create a variety of genetic devices such as: oscillators ( 13), toggle switches ( 14) and logic gates ( 15). Genetic parts libraries have been developed mostly for bacteria (especially Escherichia coli) and they have provided genetic engineers with repertoires to modulate gene expression at the transcriptional ( 2–5), post-transcriptional ( 6, 7) and translational levels ( 8–10). However, achieving predictable control of gene expression requires a detailed understanding of relevant biological processes and the availability of a sufficient number of characterized genetic parts for gene construction ( 1). coli.įorward engineering of genetic devices to carry out useful functions is at the very core of synthetic biology. Our terminator library and qTerm-Seq pipeline constitute a flexible platform enabling identification of terminator parts that can achieve transcription termination not only over a desired range but also to investigate their sequence-structure features, including for specific genetic and application contexts beyond the common in vivo systems such as E. Using qTerm-Seq, we characterize hundreds of additional strong terminators (TE > 90%) with some terminators reducing transcription read-through by up to 1000-fold in Escherichia coli. We report a rapid single-pot method to generate libraries of thousands of randomized bidirectional intrinsic terminators and a modified quantitative Term-Seq (qTerm-Seq) method to simultaneously identify terminator sequences and measure their termination efficiencies (TEs). Design principles for intrinsic terminators have been established however, additional sequence-structure studies are needed to refine parameters for termination-based genetic devices. Synthetic biology and the rational design and construction of biological devices require vast numbers of characterized biological parts, as well as reliable design tools to build increasingly complex, multigene architectures. ![]()
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